Catalogo Cell Biolabs by Fermelo Biotec

TABLE OF CONTENTS

Ordering Information & Support

Cell-Based Assays

TRANSFORMATION ADHESION MIGRATION INVASION WOUND HEALING

CELL HEALTH CELL HYPOXIA PHAGOCYTOSIS CONTRACTION ANGIOGENESIS

Stem Cell Research

IPS CELL REPROGRAMMING RETROVIRAL EXPRESSION SYSTEMS FEEDER CELLS COLONY FORMATION ASSAYS ALKALINE PHOSPHATASE ASSAYS

Viral Expression

39 ADENO-ASSOCIATED VIRUS (AAV) ADENOVIRUS LENTIVIRUS RETROVIRUS

PREMADE VIRUSES EXPRESSION SYSTEMS PURIFICATION KITS TITER KITS TRANSDUCTION KITS

microRNA Analysis

PRECURSOR CLONE COLLECTION MAMMALIAN EXPRESSION VECTORS VIRAL EXPRESSION PLASMIDS FUNCTIONAL REPORTER SYSTEM KNOCKDOWN ENHANCER

Phone 1-858-271-6500

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Fax 1-858-271-6514

TABLE OF CONTENTS

Oxidative Stress / Damage

PROTEIN OXIDATION/NITRATION LIPID PEROXIDATION DNA/RNA DAMAGE AND REPAIR HYPOXIA ASSAYS REACTIVE OXYGEN SPECIES (ROS) ASSAYS ANTIOXIDANT ASSAYS

Cell Signaling & Protein Biology

111

SMALL GTPASE / G-PROTEIN SIGNALING KINASE ASSAYS REPORTER CELLS AND REAGENTS EPITOPE TAGS PROTEIN ANALYSIS TOOLS

Metabolism Research

129

CHOLESTEROL/CHOLESTERYL ESTER ASSAYS APOLIPOPROTEIN ASSAYS AND REAGENTS FATTY ACID METABOLISM ASSAYS SERUM PROTEIN ASSAYS RENAL FUNCTION ASSAYS ALCOHOL ASSAYS

Pathogen and Toxin Assays

147

VIRUS CORE ANTIGEN ASSAYS TOXIN ASSAYS

Product Index

151

Worldwide Contacts

157

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ORDERING INFORMATION & SUPPORT Placing an Order within the U.S.

Worldwide Technical Support

Our Customer Service representatives are available Monday through Friday from 8 am to 5 pm Pacific Time.

Do you have questions about a particular product before you buy? Do you need help with a protocol?

Most orders received by 2 pm Pacific Time will be shipped the same day. We will notify you immediately of any backordered item.

Our Technical Service Scientists have been directly involved in the development and testing of our products, so you get the benefit of their hands-on experience.

We accept VISA®, MasterCard® and American Express® cards. Net 30 day terms may be offered upon credit approval.

Phone

1 858 271 6500 1 888 CBL 0505 (Toll-Free)

Phone

1 858 271 6500 1 888 CBL 0505 (US Toll-Free)

Fax

1 858 271 6514

E-mail

tech@cellbiolabs.com

Fax

1 858 271 6514

E-mail

orders@cellbiolabs.com

Pricing

Online

www.cellbiolabs.com

Mail

Attn: Customer Service 7758 Arjons Drive San Diego, CA 92126

Current U.S. prices are available online at www.cellbiolabs.com, or you may request a separate price list by e-mail. Prices are subject to change without notice. For pricing outside the U.S. please contact your local distributor, which can be found on the inside back cover of this catalog.

Placing an Order outside the U.S.

Kit Components

Please see our Worldwide Distributors section on the inside back cover of this catalog. We have a network of global distributors serving life science researchers in over 70 countries.

Because our kits are QC tested by lot, individual kit components are generally not available for purchase separately. However, certain components may be available in bulk quantities on a custom basis. For more information please inquire by sending a message to sales@cellbiolabs.com.

If you don’t see your country listed, please contact our U.S. office.

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

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CELL-BASED ASSAYS

Colony Formation Assays

Tumor Cell / Soft Agar Assays Transformation of normal cells into neoplastic cells results in a population capable of proliferating independently of internal and external signals that normally restrain growth. The soft agar colony formation assay has traditionally been used to monitor anchorageindependent growth, employing 3-4 weeks of cell growth followed by manual cell counting. We have advanced the soft agar assay to eliminate tedious manual cell counting, allow high-throughput drug screening, and enable recovery of transformed cells for downstream analysis. These advances have also allowed us to develop a unique kit for the separation of clonogenic cancer cells from normal cells in heterogeneous solid tumors.

CytoSelect™ 96-Well Cell Transformation Assay—Traditional Soft Agar Colony Formation Our CytoSelect™ 96-Well Cell Transformation Assay (Soft Agar Colony Formation) is suitable for measuring malignant transformation where no downstream analysis is required. Transformed cells cannot be recovered; however, no manual cell counting is required.

 Fast Results: 6-8 days vs. 21 days  Plate Reader Convenience: Eliminates manual counting  Versatile Format: Designed for 96-well throughput, but can be adapted for 48, 24, 12 or 6-well

With this assay, cells are incubated in a semisolid agar medium for 6-8 days, then solubilized, lysed and detected using CyQuant® GR dye in a fluorometric plate reader. Recent Product Citations 1. Eisfeld, A. et al. (2014). NRAS isoforms differentially affect downstream pathways, cell growth, and cell transformation. PNAS 111:4179-4184. 2. Gong, J. et al. (2014). Combined targeting of STAT3/NFkB/ COX-2/EP4 for effective management of pancreatic cancer. Clin. Cancer Res. 20:1259-1273. 3. Gupta, P. et al. (2013). Ocrasin targets the JNK-NFkB axis to sensitize glioma cells to TNF-alpha-induced apoptosis. Carcinogenesis 34:388-396. 4. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103. 5. Rai, V. et al. (2012). Lysophosphatidic acid targets vascular and oncogenic pathways via RAGE signaling. J. Exp. Med. 209:2339-2350. 6. Ghantous, A. et al. (2012). Inhibition of tumor promotion by parthenolide: epigenetic modulation of p21. Cancer Prev. Res. 5:1298-1309. 7. Dennis, M. et al. (2012). Snail controls the mesenchymal phenotype and drives erlotinib resistance in oral epithelial and head and neck squamous cell carcinoma cells. Carcinogenesis 147:726-732. Cell Transformation Assay Principle. Product Name

Detection

CytoSelect™ 96-Well Cell Transformation Assay (Soft Agar Colony Formation)

Fluorometric

Size

Catalog Number

1 Plate*

CBA-130

5 Plates*

CBA-130-5

*Each kit provides sufficient reagent quantities to perform 96, 48, 24, 12, or 6 tests in a 96, 48, 24, 12, or 6-well plate, respectively.

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Colony Formation Assays

CELL-BASED ASSAYS

CytoSelect™ 96-Well Cell Transformation Assays—Advanced Soft Agar with Post-Incubation Cell Recovery The CytoSelect™ 96-Well Cell Transformation Assay (Cell Recovery Compatible) provides a robust system for screening oncogenes and cell transformation inhibitors. Transformed cells may be recovered for further downstream analysis following colony formation.

Cell Transformation Assay Principle. Cell colonies form after a 6 -8 day incubation with agar matrix. Transformed cells can then be either lysed and detected with a fluorescent dye or recovered and re-plated. Product Name

 Faster Results: 6-8 days vs. 21 days  Cell Recovery: Transformed cells remain viable for further analysis  Plate Reader Convenience: Eliminates manual counting of cells  Versatile Format: Designed for 96-well throughput, but can be adapted for 48, 24, 12 or 6-well Recent Product Citations 1. Singh, R. et al. (2013). Increasing the complexity of chromatin: functionally distinct roles for replication-dependent histone H2A isoforms in cell proliferation and carcinogenesis. Nucleic Acids Res. 10.1093/nar/gkt736. (CBA-135) 2. Shukla, A. et al. (2013). Extracellular signal-regulated kinase 5: a potential therapeutic target for malignant mesotheliomas. Clin. Cancer Res. 19:2071-2083. (CBA-135) 3. Niccoli, S. et al. (2012). The Asian-American E6 variant protein of human papillomavirus 16 alone is sufficient to promote immortalization, transformation, and migration of primary human foreskin keratinocytes. J. Virol. 86:12384-12396. (CBA-135) 4. Hong, S.W. et al. (2012). Ring finger protein 149 is an E3 ubiquitin ligase active on wild-type v-Raf murine sarcoma viral oncogene homolog B1 (BRAF). J. Biol. Chem. 287:24017-24025. (CBA-135) 5. Lee, H.J. et al. (2012). Cheokine (C-X-C motif) ligand 12 is associated with gallbladder carcinoma progression and is a novel independent poor prognostic factor. Clin. Cancer Res. 18:3270-3280. (CBA-135) 6. Chapeau, E.A. et al. (2012). Ecotropic viral integration site 1 (EVI1) regulates multiple cellular processes important for cancer and is a synergistic partner for FOS proteinin invasive tumors. PNAS 109:2168-2173. (CBA-135) 7. Lim, S.K. et al. (2011). Tyrosine phosphorylation of transcriptional coactivator WW-domain binding protein 2 regulates estrogen receptor alpha function in breast cancer via the Wnt pathway. FASEB J. 25:3004-3018. (CBA-135) 8. Mathew, B. et al. (2011). The novel role of the mu opioid receptor in lung cancer progression: A laboratory investigation. Anesth. Analg. 112:558-567. (CBA-135) 9. Hirata, H. et al. (2010). Role of secreted Frizzled-related protein3 in human renal cell carcinoma. Cancer Res. 70:18961905. (CBA-135) 10. Hirata, H. et al. (2009). Wnt antagonist gene DKK2 is epigenetically silenced and inhibits renal cancer progression through apoptotic and cell cycle pathways. Clin. Cancer Res. 15:56785687. (CBA-135) Detection Colorimetric

CytoSelect™ 96-Well Cell Transformation Assay (Cell Recovery Compatible) Fluorometric

CytoSelect™ 384-Well Cell Transformation Assay**

Fluorometric

Size

Catalog Number

1 Plate*

CBA-135

5 Plates*

CBA-135-5

1 Plate*

CBA-140

5 Plates*

CBA-140-5

1 Plate***

CBA-145

5 Plates***

CBA-145-5

*Each kit provides sufficient reagent quantities to perform 96, 48, 24, 12, or 6 tests in a 96, 48, 24, 12, or 6-well plate, respectively. **The 384-well kit does not allow for cell recovery due to small well size. ***Each kit provides sufficient reagents for one or five 384-well plates respectively.

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CELL-BASED ASSAYS

Colony Formation Assays

CytoSelect™ 96-Well In Vitro Tumor Sensitivity Assay The CytoSelect™ In Vitro Tumor Sensitivity Assay provides a stringent, anchorage-independent model for chemosensitivity testing and possible anticancer drug screening. The assay uses a soft agar matrix to promote the colony formation of neoplastic cells in about a week. Cells are quantified using a standard ELISA plate reader.  Fast Results: 6-8 days  In Vivo Simulation: Resembles a threedimensional cell environment  Plate Reader Convenience: Eliminates manual counting

Tumor Sensitivity Assay Principle.

Inhibition of HeLa Cell Anchorage-Independent Growth by Taxol. HeLa cells were cultured for 7 days in the absence (top) or presence (bottom) of 1 nM Taxol according to the assay protocol.

Product Name

Detection

CytoSelect™ 96-Well In Vitro Tumor Sensitivity Assay

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Recent Product Citations 1. Bard-Chapeau, E. et al. (2013). EVI1 oncoprotein interacts with a large and complex network of proteins and integrates signals through protein phosphorylation. PNAS 110:E2885-E2894. 2. Takezawa, K. et al. (2012). HER2 amplification: a potential mechanism of acquired resistance to EGFR inhibition in EGFRmutant lung cancers that lack the second-site EGFRT790M mutation. Cancer Discovery 2:922-933. 3. Li,C. et al. (2012). The root bark of Paeonia moutan is a potential anticancer agent in human oral squamous cell carcinoma cells. Anticancer Res. 32:2625-2630. 4. Itamochi, H. et al. (2011). Inhibiting the mTOR pathway synergistically enhances cytotoxicity in ovarian cancer cells induced by etoposide through upregulation of c-Jun. Clin. Cancer Res. 17:4742-4750. 5. Kang, D.W. et al. (2010). Phospholipase D1 drives a positive feedback loop to reinforce the Wnt/ß-catenin/TCF signaling axis. Cancer Res. 70:4233-4242.

Colorimetric

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Size

Catalog Number

96 Assays

CBA-150

5 x 96 Assays

CBA-150-5

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Colony Formation Assays

CELL-BASED ASSAYS

CytoSelect™ Clonogenic Tumor Cell Isolation Kit Clean separation of clonogenic tumor cells from normal cells is critical for proper analysis of disease state progression. Due to the heterogeneity of many tumors, however, isolation of homogenous tumor cell populations can be difficult. The CytoSelect™ Clonogenic Tumor Cell Isolation Kit uses a proprietary semisolid agar medium to facilitate colony formation by cells from solid tumors. Colonies are grown in either a 6-well plate or a 35 mm dish. The colonies are then isolated away from single cells by size filtration.

 Efficient: Easily eliminates single cells from clonogenic tumor cell population  Versatile: In addition to solid tumors, has potential use in isolating tumor stem cells

Clonogenic Colony Formation, Isolation and Re-plating. A: Clonogenic colony formation (red arrows) and single cells (black arrows) after 7 day incubation. B: Isolation of clonogenic colonies from single cells. C: Re-plated clonogenic colonies after 3 days (no trypsinization). D: Re-plated clonogenic colonies 1 day after trypsinization.

Clonogenic Tumor Cell Isolation Procedure.

Product Name CytoSelect™ Clonogenic Tumor Cell Isolation Kit

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Size

Catalog Number

5 Preps

CBA-155

25 Preps

CBA-155-5

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CELL-BASED ASSAYS

Cell Adhesion

Cell Adhesion Assays Cell adhesion is a complex mechanism involved in a variety of processes including cell migration/invasion, embryogenesis, wound healing and tissue remodeling. Cells can adhere to the ECM, forming complexes with cytoskeletal components, or to the endothelium. Our CytoSelect™ Cell Adhesion Assays quantify adhesion of cells using a microplate reader or fluorometer; no manual cell counting is required.

CytoSelect™ Leukocyte Endothelium Adhesion Assays The CytoSelect™ Leukocyte Endothelium Adhesion Assays provide a robust system for the quantitative determination of interactions between leukocytes and endothelium. Adherent cells can be quantified on a fluorescence plate reader. Recent Product Citations 1. Liu, G. et al. (2012). ICAM-1-activated Src and eNOS signaling increase endothelial cell surface PECAM-1 adhesivity and neutrophil transmigration. Blood 120:1942-1952. (CBA-210) 2. Fang, Y. et al. (2012). Site-specific microRNA-92a regulation of Kruppel-like factors 4 and 2 in atherosusceptible endothelium. Arterioscler. Thromb. Vasc. Biol. 32:979-987. (CBA-210) 3. Curatola, A.M. et al. (2012). Dehydroepiandrosterone (DHEA) inhibition of monocyte binding by vascular endothelium is associated with sialylation of neural cell adhesion of neural cell adhesion molecule. Repr. Sciences 19:86-91. (CBA-210) 4. Wang, Y.L. et al. (2011). Innate immune function of the adherens junction protein p120-catenin in endothelial response to endotoxin. J. Immunol. 186:3180-3187. (CBA-210) 5. Ziemann, A. et al. (2013). CRN2 enhances the invasiveness of glioblastoma cells. Neuro Oncology 10.1093/neuonc/ nos388. (CBA-215) 6. Hernandez, L. et al. (2010). Activation of NF-kB signaling by inhibitor of NF-kB Kinase ß increases aggressiveness of ovarian cancer. Cancer Res. 70:4005-4014. (CBA-215) Product Name CytoSelect™ Leukocyte-Endothelium Adhesion Assay

CytoSelect™ Leukocyte-endothelium Adhesion Assay Principle. Detection

Size

Catalog Number

Fluorometric

96 Assays

CBA-210

CytoSelect™ Leukocyte-Epithelium Adhesion Assay

Fluorometric

96 Assays

CBA-211

CytoSelect™ Tumor-Endothelium Adhesion Assay

Fluorometric

96 Assays

CBA-215

CytoSelect™ Microfluidic Biochips CytoSelect™ 8-Channel Microfluidic Biochips provide an environment that closely mimics in vivo shear stresses, resulting in more physiologically relevant cell adhesion data. The Biochips are self-contained units containing 8 channels with inlet/outlet ports at both ends of each channel. After adding cell samples, a syringe pump set to the proper flow rate applies the appropriate shear stress to the channel.

Product Name

Detection

CytoSelect™ 8-Channel ECM Microfluidic Biochips

Microscopy

CytoSelect™ 8-Channel Endothelial Microfluidic Biochips

Microscopy

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Size

Catalog Number

2 Chips

CBA-003

10 Chips

CBA-003-5

2 Chips

CBA-004

10 Chips

CBA-004-5

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Cell Adhesion

CELL-BASED ASSAYS

CytoSelect™ ECM Cell Adhesion Assays The CytoSelect™ ECM Cell Adhesion Assays provide a quantitative method to measure cell adhesion. The 48well plate is precoated with your choice of substrate. Adherent cells attach, while non-adherent cells are washed away. Adherent cells can be quantified on a standard plate reader or fluorometer.  Quantitative: Measure results in a colorimetric or fluorescence plate reader  Flexible: Uniform substrate layer of your choice of Collagen I, Collagen IV, Fibrinogen, Fibronectin, or Laminin; or choose the ECM array which contains all 5 ECM proteins

BSA Vitronectin Laminin Fibronectin Collagen IV Collagen I MDA-231 HT-1080 HEK293 CytoSelect™ 48-well Cell Adhesion Assay. Serum starved cells from three different cell lines were allowed to attach to the ECMcoated plate for 1 hr at 100,000 cells/well. Adherent cells were stained according to the assay protocol. Product Name

Detection

CytoSelect™ 48-Well Cell Adhesion Assay, ECM Array (Contains one row each of Collagen I, Collagen IV, Fibrinogen, Fibronectin, and Laminin)

CytoSelect™ 48-Well Cell Adhesion Assay, Collagen I

CytoSelect™ 48-Well Cell Adhesion Assay, Collagen IV

CytoSelect™ 48-Well Cell Adhesion Assay, Fibrinogen

CytoSelect™ 48-Well Cell Adhesion Assay, Fibronectin

CytoSelect™ 48-Well Cell Adhesion Assay, Laminin

Recent Product Citations 1. Cervera, A.M. et al. (2008). Cells silenced for SDHB expression display characteristic features of the tumor phenotype. Cancer Res. 68:4058-4067. (CBA-050 and CBA-070) 2. Lee, J. et al. (2013). Selective inhibition of prostaglandin E2 receptors EP2 and EP4 inhibits adhesion of human endometriotic epithelial and stromal cells through suppression of integrinmediated mechanisms. Biol. Reprod. 88:77. (CBA-057) 3. Miao, H. et al. (2008). Gene expression and functional studies of the optic nerve head astrocyte transcriptome from normal African Americans and Caucasian Americans donors. PLoS One 3(8):E2847. (CBA-060) 4. Tanoury, Z. et al. (2014). Genes involved in cell adhesion and signaling: a new repertoire of retinoic acid receptor target genes in mouse embryonic fibroblasts. J. Cell Sci. 127:521533. (CBA-070) 5. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103. (CBA-070) 6. Mei, J. et al. (2012). Inhibition of IDO1 suppresses cyclooxygenase-2 and matrix metalloporeinase-9 expression and decreases proliferation, adhesion and invasion of endometrial stromal cells. Mol. Human Reprod. 10.1093/molehr/gas021. (CBA-070) 7. Kandalam, V. et al. (2011). Lack of tissue inhibitor of metalloproteinases 2 leads to exacerbated left ventricular dysfunction and adverse extracellular matrix remodeling in response to biomechanical stress. Circulation 124:2094-2105. (CBA-070) 8. Tsukamoto, H. et al. (2010). Evaluation of anticancer activities of benzo[c]phenanthridine alkaloid sanguinarine in oral squamous cell carcinoma cell line. Anticancer Res. 31:28412846. (CBA-070) 9. Kim, S.W. et al. (2013). Cardiac stem cells with electrical stimulation improve ischaemic heart function through regulation of connective tissue growth factor and miR-378. Cardiovasc. Res. 10.1093/cvr/cvt192. (CBA-071) Size

Catalog Number

48 Assays

CBA-070

5 x 48 Assays

CBA-070-5

48 Assays

CBA-071

5 x 48 Assays

CBA-071-5

Colorimetric

48 Assays

CBA-052

Fluorometric

48 Assays

CBA-053

Colorimetric

48 Assays

CBA-060

Fluorometric

48 Assays

CBA-061

Colorimetric

48 Assays

CBA-058

Fluorometric

48 Assays

CBA-059

Colorimetric

48 Assays

CBA-050

Fluorometric

48 Assays

CBA-051

Colorimetric

48 Assays

CBA-056

Fluorometric

48 Assays

CBA-057

Colorimetric

Fluorometric

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CELL-BASED ASSAYS

Cell Migration / Invasion

Cell Migration & Invasion Assays Cell migration and invasion are highly integrated, multi-step processes and play important roles in the progression of various diseases including cancer, atherosclerosis and arthritis. Our cell migration assays are provided in two formats: 2-Dimensional Gap Closure and Boyden Chamber. Each format has its own advantages and applications. Use the information below to help choose the best format for your cell migration experimental goals. Cell invasion assays are provided in the Boyden Chamber format.

Cell Migration Format Selection Guide 2D Gap Closure Assays (p. 13)

Boyden Chamber Assays (p. 14-19)

Qualitative or Quantitative

Quantitative

Endpoint or Real Time

Endpoint

Microscopy

Plate Reader

Yes

Relative Sensitivity

Good

Fair

Adaptability to Automation

Good

Poor

Universal

Choose pore size based on cell size

Type of Analysis Detection Time Detection Method Chemoattractant Gradient

Cell Compatibility

Assay Principle for the Radiusâ&#x201E;˘ 2D Gap Closure Assays.

Example of Boyden Chamber Assay Principle.

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Cell Migration / Invasion

CELL-BASED ASSAYS

Radius™ Cell Migration Assays (2D Gap Closure) Radius™ Cell Migration Assays provide a unique alternative to the traditional Boyden Chamber migration assay. Radius assays allow you to measure cell migration at endpoint or in real time, and are ideal for time course migration studies. Radius™ Cell Migration Assays use a cell culture plate containing a proprietary, carefully-defined biocompatible hydrogel (Radius™ gel) spot centralized at the bottom of each well. Cells seeded in the well will attach everywhere except on the Radius gel spot, creating a cell-free zone. Once cells attach, the Radius gel is removed and migration of cells across the cell-free zone begins. The gel removal step allows synchronization of a zero time point to facilitate wellto-well comparisons. With Radius™ Cell Migration Assays, there are no cell culture inserts; so you don’t need to worry about which pore size to choose for your cell type. Any adherent cell may be used in the assay. Radius assays are supplied in 24-well, 96-well and 384-well formats. In addition, the 24-well assays are provided with your choice of coatings for proper cell attachment:     

Uncoated Collagen I-coated Fibronectin-coated Laminin-coated ECM Array with 6 wells of each of the above (uncoated, Collagen I, Fibronectin, Laminin); ideal if you are unsure which ECM protein may provide the best cell attachment Product Name

Example Results using 2D Gap Closure Assay. Recent Product Citations 1. Sun, J. et al. (2013). Targeting the metastasis suppressor, NDRG1, using novel iron chelators: regulation of stress fibermediated tumor cell migration via modulation of the ROCK1/ pMLC2 signaling pathway. Mol. Pharmacol. 83:454-469. (CBA125) 2. Apostolos, K. et al. (2013). Increased susceptibility of melaninconcentrating hormone-deficient mice to infection with Salmonella enterica serovar Typhimurium. Infect. Immun. 81:166-172. (CBA125) 3. Smith, K. et al. (2012). Human family with sequence similarity 60 member A (FAM60A) protein: a new subunit of the Sin3 deacetylase complex. Mol. Cell Proteomics 11:1815-1828. (CBA-125) 4. Young, S. et al. (2012). Rapid protein kinase D1 signaling promotes migration of intestinal epithelial cells. Am. J. Physiol. Gastrointest. Liver Physiol. 303:G356-G366. (CBA-125) 5. Larive, R.M. et al. (2012). The Ras-like protein R-Ras2/TC21 is important for proper mammary gland development. Mol. Biol. Cell 23:2373-2387. (CBA-125) 6. Wong, B. et al. (2013). Adrenomedullin enhances invasion of human extravillous cytotrophoblast-derived cell lines by regulation of urokinase plasminogen activator expression and S-nitrosylation. Biol. Reprod. 88:34. (CBA-126) 7. Ichikawa, A. et al. (2013). CXCL10-CXCR3 enhances the development of neutrophil-mediated fulminant lung injury of viral and nonviral origin. Am. J. Respir. Crit. Care Med. 187:65-77. (CBA-126) 8. Coulouarn, C. et al. (2012). Hepatocyte-stellate cell cross-talk in the liver engenders a permissive inflammatory microenvironment that drives progression in hepatocellular carcinoma. Cancer Res. 72:2533-2542. (CBA-126) 9. Alcolea, S. et al. (2012). Interaction between head and neck squamous cell carcinoma cells and fibroblasts in the biosynthesis of PGE2. J. Lipid Res. 53:630-642. (CBA-126) Detection

Size

Catalog Number

24 Assays

CBA-125

5 x 24 Assays

CBA-125-5

Radius™ 24-Well Cell Migration Assay

Microscopy

Radius™ 24-Well Cell Migration Assay (Collagen I Coated)

Microscopy

24 Assays

CBA-125-COL

Radius™ 24-Well Cell Migration Assay (Fibronectin Coated)

Microscopy

24 Assays

CBA-125-FN

Radius™ 24-Well Cell Migration Assay (Laminin Coated)

Microscopy

24 Assays

CBA-125-LN

Radius™ 24-Well Cell Migration Assay (ECM Array Coated)

Microscopy

24 Assays

CBA-125-ECM

96 Assays

CBA-126

Radius™ 96-Well Cell Migration Assay

Microscopy

5 x 96 Assays

CBA-126-5

384 Assays

CBA-127

Radius™ 384-Well Cell Migration Assay

Microscopy

5 x 384 Assays

CBA-127-5

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CELL-BASED ASSAYS

Cell Migration / Invasion

CytoSelect™ Cell Migration and Invasion Assays (Boyden Chamber) The Boyden Chamber has been extensively used and widely published as a tool for measuring cell migration and cell invasion in vitro. Our CytoSelect™ Cell Migration and Invasion Assays use this well-cited method to quantify cell migration and invasion with no manual cell counting required. Migratory or invasive cells are quantified using a colorimetric or fluorometric plate reader. Cell migration may take on various forms and behaviors depending on the type and location of cells. Such subclasses of cell migration include chemotaxis, haptotaxis, and transmigration. Use the chart below to compare the various subclasses of cell migration as well as cell invasion, which will help you choose the assay best suited to your experimental goals.

Typical Well Setup for Boyden Chamber Assay.

Boyden Chamber Assay Selection Guide Assay

Chemotaxis (p. 15)

Haptotaxis (p. 16)

Definition

Migration of cells toward a chemoattractant (chemical signal) in the cell’s surrounding environment

Migration of cells along a gradient of cellular adhesion sites or extracellular matrixbound chemoattractants

Migration of cells through the Transmigration vascular endothelium toward a (p. 17) chemoattractant

Invasion (p. 18-19)

Movement of cells through the 3D extracellular matrix into neighboring tissues; includes ECM degradation and proteolysis

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Cell Types

Pore Size

Insert Coating

Assay Formats

Neutrophils Leukocytes

3 µm

None

24-Well 96-Well

Lymphocytes Monocytes Macrophages

5 µm

None

24-Well 96-Well

Fibroblasts Endothelial Cells Epithelial Cells Tumor Cells

8 µm

None

24-Well 96-Well

Astrocytes 12 µm Slow-moving Cells

None

24-Well

Collagen I (bottom)

24-Well

Fibronectin (bottom)

24-Well

Fibroblasts Endothelial Cells Epithelial Cells

8 µm

Leukocytes

3 µm

None

24-Well

Tumor Cells

8 µm

None

24-Well

ECM Matrix (top)

24-Well 96-Well

Collagen I (top)

24-Well 96-Well

Laminin I (top)

24-Well 96-Well

Fibroblasts Endothelial Cells Epithelial Cells Tumor Cells

8 µm

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Cell Migration / Invasion

CELL-BASED ASSAYS

CytoSelect™ Cell Migration Assays—Chemotaxis CytoSelect™ Cell Migration Assays are ideal for measuring chemotaxis. The kits utilize polycarbonate membrane inserts in 24-well or 96-well plates. Inserts are available with 4 different pore sizes to accommodate a variety of cell types.  Fast Results: Visualize chemotaxis in less than 6 hours with most cell types  Flexible: Bottoms of membrane inserts are uncoated to allow use with any chemoattractant  Higher Throughput: 96-well format available for fluorescence plate readers

0% FBS

10% FBS

Migration of Human Fibrosarcoma HT-1080 Cells. Cells were seeded at 30,000 cells per well of a 24-well plate and allowed to migrate toward 10% FBS for 4 hours. Migratory cells were stained (above) and quantified in a fluorescence plate reader (data not shown).

Product Name

Recent Product Citations 1. Al-Wadei, M.H. et al. (2013). Gamma-amino butyric acid (GABA) prevents the induction of nicotinic receptor-reuglated signaling by chronic ethanol in pancreatic cancer cells and normal duct epithelia. Cancer Prev. Res. 6:139-148. (CBA-100) 2. Chen, Z. et al. (2012). The iron chelators Cp44mT and DFO inhibit TGF-ß-induced epithelial-mesenchymal transition via upregulation of N-Myc downstream-regulated gene 1 (NDRG1). J. Biol. Chem. 287:17016-17028. (CBA-100) 3. Izhak, L. et al. (2010). Predominant expression of CCL2 at the tumor site of prostate cancer patients directs a selective loss of immunological tolerance to CCL2 that could be amplified in a beneficial manner. J. Immunol. 184:1092-1101. (CBA-101) 4. Kiss, J. et al. (2012). Loss of the oxygen sensor PHD3 enhances the innate immune response to abdominal sepsis. J. Immunol. 189:1955-1965. (CBA-102) 5. Verma, S. et al. (2011). Selenoprotein K knockout mice exhibit deficient calcium flux in immune cells and impaired immune responses. J. Immunol. 186:2127-2137. (CBA-103) 6. Li, X. et al. (2011). Kaposi’s sarcoma-associated Herpesvirusencoded latency-associated nuclear antigen recduces interleukin8 expression in endothelial cells and impairs neutrophil chemotaxis by degrading nuclear p65. J. Virol. 85:8606-8615. (CBA-104) 7. Jun, H.S. et al. (2012). Glucose-6-phosphatase-ß, implicated in a congenital neutropenia syndrome, is essential for macrophage energy homeostasis and functionality. Blood 119:4047-4055. (CBA-105) 8. Rosenblum, S. et al. (2012). Timing of intra-arterial neural stem cell transplantation after hypoxia-ischemia influences cell engraftment, survival, and differentiation. Stroke 43:1624-1631. (CBA106) 9. Aftab, B.T. et al. (2011). Itraconazole inhibits angiogenesis and tumor growth in non-small cell lung cancer. Cancer Res. 71:67646772. (CBA-106)

Pore Size

Detection

Size

Catalog Number

3 µm

Fluorometric

12 Assays

CBA-103

5 x 12 Assays

CBA-103-5

5 µm

Fluorometric

12 Assays

CBA-102

5 x 12 Assays

CBA-102-5

12 Assays

CBA-100

5 x 12 Assays

CBA-100-5

12 Assays

CBA-101

5 x 12 Assays

CBA-101-5

Colorimetric

12 Assays

CBA-107

Fluorometric

12 Assays

CBA-108

96 Assays

CBA-104

5 x 96 Assays

CBA-104-5

96 Assays

CBA-105

5 x 96 Assays

CBA-105-5

96 Assays

CBA-106

5 x 96 Assays

CBA-106-5

Colorimetric

CytoSelect™ 24-Well Cell Migration Assay 8 µm

Fluorometric

12 µm

CytoSelect™ 96-Well Cell Migration Assay

3 µm

Fluorometric

5 µm

Fluorometric

8 µm

Fluorometric

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CELL-BASED ASSAYS

Cell Migration / Invasion

CytoSelect™ Cell Migration Assays—Haptotaxis Haptotaxis describes the migration of cells toward a gradient of immobilized extracellular matrix. The CytoSelect™ Cell Haptotaxis Assays are ideal for determining the migratory properties of cells. The kits utilize polycarbonate membrane inserts with an 8 µm pore size in a 24-well plate. The undersides of the inserts are coated with either Collagen or Fibronectin. The 8 µm pore size in the membrane inserts is ideal for epithelial cells, endothelial cells, fibroblasts, and other cells of similar size. The membrane serves as a barrier that allows discrimination of migratory cells from non-migratory cells.  Fast Results: Visualize cell haptotaxis in less than 6 hours with most cell types  Convenient: Membrane inserts pre-coated on the underside with either Collagen I or Fibronectin  Versatile: Useful with a variety of cell types including epithelial cells, endothelial cells, and fibroblasts*

Assay Principle for the CytoSelect™ Cell Haptotaxis Assay.

BSA

Collagen I

Fibronectin

*For leukocyte migration a 3 µm pore size is recommended. See our CytoSelect™ Chemotaxis Assays (previous page) or the CytoSelect™ Leukocyte Transmigration Assay (next page).

0% FBS Recent Product Citations 1. Herrera, I. et al. (2013). Matrix metalloproteinase (MMP)-1 induces lung alveolar epithelial cell migration and proliferation, protects from apoptosis, and represses mitochondrial oxygen consumption. J. Biol. Chem. 288:25964-25975. (CBA-100-COL) 2. Niccoli, S. et al. (2012). The Asian-American E6 variant protein of human papillomavirus 16 alone is sufficient to promote immortalization, transformation, and migration of primary human foreskin keratinocytes. J. Virol. 86:12384-12396. (CBA-110-COL) 3. Kamiya, K. et al. (2007). Protein Kinase C delta activated adhesion regulates vascular smooth muscle cell migration. J. Surg. Res. 141:91-96. (CBA-100-COL)

Product Name CytoSelect™ 24-Well Cell Haptotaxis Assay, Collagen I-coated

CytoSelect™ 24-Well Cell Haptotaxis Assay, Fibronectin-coated

Phone 1-858-271-6500

0.5% FBS

CytoSelect™ 24-well Cell Haptotaxis Assay. MDA-231 cells were seeded at 150,000 cells/well and allowed to migrate toward FBS for 4 hrs. Migratory cells, found on the bottom of the migration membrane, were stained according to the assay protocol.

Detection

Size

Catalog Number

Colorimetric

12 Assays

CBA-100-COL

Fluorometric

12 Assays

CBA-101-COL

Colorimetric

12 Assays

CBA-100-FN

Fluorometric

12 Assays

CBA-101-FN

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Cell Migration / Invasion

CELL-BASED ASSAYS

CytoSelect™ Cell Migration Assays—Transmigration Cancer cell transmigration, particularly extravasation, is an important step in cancer metastasis. The CytoSelect™ Cell Transmigration Assays provide a robust system for the quantitation of transmigrations and interactions between endothelium and cancer cells. Migratory cells are quantified via fluorometer.

The Leukocyte Adhesion and Transmigration Cascade. 200

RFU

160 120 80 40 0 0

10,000

20,000

30,000

40,000

50,000

Cell Number Quantitation of Human Monocytic THP-1. LeukoTracker™ labeled THP-1 cells were titrated in 1X PBS, then lysed with 2X lysis buffer. Fluorescence was quantified as described in the assay protocol. Recent Product Citations 1. Fava, G. et al. (2008). Leptin enhances cholangiocarcinoma cell growth. Cancer Res. 68:6752-6761. (CBA-212) 2. Park, G.B. et al. (2014). The Epstein-Barr Virus causes epithelial -mesenchymal transition in human corneal epithelial cells via Syk/Src and Akt/ERK signaling pathways. Invest. Ophthalmol. Vis. Sci. 55:1770-1779. (CBA-216) 3. Xu, Z. et al. (2010). Role of pancreatic stellate cells in pancreatic cancer metastasis. Am. J. Pathol. 177:2585-2596. (CBA-216)

Assay Principle for the CytoSelect™ Leukocyte Transmigration Assay.

Product Name

Pore Size

Detection

Size

Catalog Number

CytoSelect™ Leukocyte Transmigration Assay

3 µm

Fluorometric

24 Assays

CBA-212

CytoSelect™ Tumor Transendothelial Migration Assay

8 µm

Fluorometric

24 Assays

CBA-216

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CELL-BASED ASSAYS

Cell Migration / Invasion

CytoSelect™ Cell Invasion Assays Tumor cell invasion into surrounding normal tissue contributes to the morbidity of cancers. The CytoSelect™ Cell Invasion Assays use precoated inserts to assay invasive properties of tumor cells in 24-well or 96-well plates. The coated layer serves to distinguish invasive cells from non-invasive cells. Plates are precoated with either basement membrane matrix (from EHS mouse sarcoma cells), Collagen I or Laminin I.

 Quantitative: Measure results in a colorimetric or fluorescence plate reader  Flexible: Uniform protein matrix layer of your choice of basement membrane (from mouse tumor cells), Collagen I, or Laminin I  Versatile: Characterize both the invasive and migratory properties of your cells with a Cell Migration / Invasion Combo Kit (next page)

CytoSelect™ Cell Invasion Assay Principle. Recent Product Citations 1. Gradilone, S. et al. (2013). HDAC6 inhibition restores ciliary expression and decreases tumor growth. Cancer Res. 73:22592270. (CBA-110) 2. Nam, H. et al. (2013). Antitumor activity of saracatinib (AZD0530), a c-Src/Abl kincase inhibitor, alone or in combination with chemotherapeutic agents in gastric cancer. Mol. Cancer Ther. 12:16-26. (CBA-110) 3. DiNatale, B. et al. (2012). Ah receptor antagonism represses head and neck tumor cell aggressive phenotype. Mol. Cancer Res. 10:1369-1379. (CBA-110) 4. Citterio, C. et al (2012). The Rho exchange factors Vav2 and Vav3 control a lung metastasis-specific transcriptional program in breast cancer cells. Sci. Signal. 5:ra71. (CBA-110) 5. Nakayama, K. et al. (2013). cAMP-response element-binding protein (CREB) and NFkB transcription factors are activated during prolonged hypoxia and cooperatively regulate the induction of matrix metalloproteinase MMP1. J. Biol. Chem. 288:258422595. (CBA-112) 6. Desai, S.D. et al. (2012). ISG15 disrupts cytoskeletal architecture and promotes motility in human breast cancer cells. Exp. Biol. Med. 237:38-49. (CBA-112) 7. Ziemann, A. et al. (2013). CRN2 enhances the invasiveness of gliobastoma cells. Neuro Oncology 10.1093/neuonc/nos388. (CBA-112-COL) 8. Liu, X. et al. (2013). Antiproliferative, antiinvasive, and proapoptotic activity of folate receptor alpha-targeted liposomal doxorubicin in nonfunctional pituitary adenoma cells. Endocrinology 154:1414-1423. (CBA-112-COL)

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Human Fibrosarcoma HT-1080 Laminin I Cell Invasion. HT1080 and NIH3T3 (negative control) were seeded at 200,000 cells/ well and allowed to invade toward FBS for 24 hrs. Invasive cells on the membrane bottom were stained (top and center) and quantified at OD 560nm after extraction (data not shown).

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Cell Migration / Invasion

CELL-BASED ASSAYS

CytoSelect™ Cell Invasion Assays, continued 2

HT-1080

1.6 OD 560nm

NIH3T3

1.2 0.8 0.4 0 NIH3T3

HT-1080

HT-1080 + Cytochalasin D

Effects of Cytochalasin D on Invading Cells using the CytoSelect™ 24-well Cell Invasion Assay (CBA-110). HT-1080 and NIH3T3 cells (negative control) were seeded at 300,000 cells/well and allowed to invade toward 10% FBS for 24 hrs, in the presence or absence of 2 µM Cytochalasin D. Invasive cells, on the bottom of the invasion membrane, were stained (left) and then quantified at OD 560 nm after extraction using a standard plate reader (right). Product Name

Detection

Size

Catalog Number

Colorimetric

12 Assays

CBA-110

Fluorometric

12 Assays

CBA-111

Colorimetric

12 Assays

CBA-110-COL

Fluorometric

12 Assays

CBA-111-COL

Colorimetric

12 Assays

CBA-110-LN

Fluorometric

12 Assays

CBA-111-LN

CytoSelect™ 96-Well Cell Invasion Assay, Basement Membrane

Fluorometric

96 Assays

CBA-112

CytoSelect™ 96-Well Cell Invasion Assay, Collagen I

Fluorometric

96 Assays

CBA-112-COL

CytoSelect™ 96-Well Cell Invasion Assay, Laminin I

Fluorometric

96 Assays

CBA-112-LN

CytoSelect™ 24-Well Cell Invasion Assay, Basement Membrane

CytoSelect™ 24-Well Cell Invasion Assay, Collagen I

CytoSelect™ 24-Well Cell Invasion Assay, Laminin I

CytoSelect™ Cell Migration / Invasion Assay Combo Kits Our CytoSelect™ Cell Migration / Invasion Assay Combo Kits allow you to characterize both the migratory and invasive properties of your cells. Each 24-well combo kit provides sufficient reagents to perform 12 migration plus 12 invasion assays, while the 96-well combo kit allows you to perform 96 migration plus 96 invasion assays. The invasion plate provided contains basement membrane-coated inserts. Recent Product Citations 1. Majid, S. et al. (2013). miRNA-34b inhibits prostate cancer through demethylation, active chromatin modification, and AKR pathways. Clin. Cancer Res. 19:73-84. (CBA-100-C) 2. Shin, S.Y. et al. (2012). Transcriptional regulation of the interleukin-11 gene by oncogenic Ras. Carcinogenesis 10.1093/carcin/bgs297. (CBA-100-C) 3. Gobeil, S. et al. (2008). A genome-wide shRNA screen identifies GAS1 as a novel melanoma metastasis suppressor gene. Genes Dev. 22(21):2932-2940. (CBA-101-C) 4. Axlund, S.D. et al. (2010). HOXC8 inhibits androgen receptor signaling in human prostate cancer cells by inhibiting SRC-3 recruitment to direct androgen target genes. Mol. Cancer Res. 8:1643-1655. (CBA-106-C) Product Name

Pore Size

CytoSelect™ 24-Well Cell Migration / Invasion Combo Kit

8 µm

CytoSelect™ 96-Well Cell Migration / Invasion Combo Kit

8 µm

Detection

Size

Catalog Number

Colorimetric

2 x 12 Assays

CBA-100-C

Fluorometric

2 x 12 Assays

CBA-101-C

Fluorometric

2 x 96 Assays

CBA-106-C

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CELL-BASED ASSAYS

Cell Migration / Invasion

CytoSelect™ 24-Well Wound Healing / Cell Migration Assay Compared to traditional scratch assays, our CytoSelect™ 24-Well Wound Healing Assay provides a more consistent method to measure cell migration across a “wound field” gap in vitro. Proprietary treated inserts generate a consistently defined 0.9mm gap between the cells. Cells can then be treated and monitored for proliferation or migration across the wound field by imaging samples at fixed time points or time-lapse microscopy.

 Highly Accurate: More consistent results well-towell compared to homemade scratch assays  Versatile: Measure cell migration, cell proliferation, and wound closure  Inert Material: No residues from inserts to impede cell migration or proliferation

Percent Wound Closure

Recent Product Citations 1. Cui, X.G. et al. (2013). Response gene to complement 32 deficiency causes impaired placental angiogenesis in mice. Cardiovasc. Res. 10.1093/cvr/cvt121. 2. Gutschner, T. et al. (2013). The noncoding RNA MALAT1 is a critical regulator of the metastasis phenotype of lung cancer cells. Cancer Res. 73:1180-1189. 3. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103. 4. Mao, R. et al. (2012). Circadian gating of epithelial-tomesenchymal transition in breast cancer cells via melatoninregulation of GSK3ß. Mol Endrocrinol. 26:1808-1820. 5. Hirata, H. et al. (2012). MicroRNA-1826 targets VEGFC, betacatenin (CTNNB1) and MEK1 (MAP2K1) in human bladder cancer. Carcinogenesis 33:41-48.

50%

100%

CytoSelect™ 24-well Wound Healing Assay Principle.

Wound Closure of STO Cells. STO cells (mouse MEF) were cultured in the provided plate with inserts in place for 24 hours until a monolayer formed. Inserts were then removed to begin the assay. Cells were monitored at various time points and stained according to the assay protocol.

Product Name

Detection

CytoSelect™ 24-Well Wound Healing Assay

Microscopy

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

24 Assays

CBA-120

5 x 24 Assays

CBA-120-5

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Cell Co-Culture

CELL-BASED ASSAYS

CytoSelect™ 24-Well Cell Co-Culture System The culture of two cell lines together is advantageous for studying a variety of applications: 

Cell-cell interactions



Cell activation



Cellular differentiation



Maintaining stem cell pluripotency



Various effects of secreted factors from one cell type on a second cell type

Traditional methods of co-culture usually involve one of the following methods: 1. One cell type is cultured to form a monolayer, followed by seeding of a second cell type directly over the monolayer. This is a common method when feeder cells are used to maintain stem cells in an undifferentiated state. However, it is not useful when studying the effects of one cell on the other because the first cell is obscured from view by the second. 2. A Boyden Chamber (see page 12 for details) is used to culture one cell type above the membrane and a second cell type below the membrane. This system allows a separation between the two cells, but does not allow for direct cell-tocell contact which may reduce its efficacy for certain applications. The CytoSelect™ Cell Co-Culture System provides a unique platform for direct contact between two cell types in one well. A proprietary molded plastic insert creates a cell-free zone in the center of a 24-well cell culture-treated plate. The first cell type is seeded in the area around the insert. Once the cells form a monolayer, the insert is removed and the second cell is seeded.

Protocol for the CytoSelect™ 24-Well Co-Culture System. Cell type #1 is seeded with the Co-Culture insert in place. After cells form a monolayer, the insert is removed and cell type #2 is seeded.

Product Name

Detection

Size

Catalog Number

CytoSelect™ 24-Well Cell Co-Culture System

Microscopy

24 Assays

CBA-160

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CELL-BASED ASSAYS

Cell Health

CytoSelect™ Cell Viability and Cytotoxicity Assay (Live/Dead) Cell viability characteristics include cellular metabolic activity and cell membrane integrity. Our CytoSelect™ Cell Viability and Cytotoxicity Assay provides both a colorimetric and fluorometric format for monitoring cell viability via metabolic activity. Live cells are detected with MTT (colorimetric detection) or Calcein AM (fluorometric); dead cells are detected with EthD1 reagent (fluorometric). All 3 detection reagents are included, as well as Saponin, a cell death initiator. Cells may be treated with compounds or agents that affect cell viability. This kit is suitable for eukaryotic cells, not bacteria or yeast.  Versatile: Detect by microscopy, colorimetric or fluorescence plate reader, or flow cytometry  Quantitative: Measure live and dead cells on a fluorescence plate reader; live cells may also be quantified on a standard microplate reader Recent Product Citation Kim, E.Y. et al. (2012). Sustained activation of N-methyl-Daspartate receptors in podocytes leads to oxidative stress, mobilization of transient receptor potential canonical 6 channels, nuclear factor of activated T cells activation, and apoptotic cell death. Mol. Pharmacol. 82:728-737. Product Name CytoSelect™ Cell Viability and Cytotoxicity Assay Kit

Viability of Human Foreskin Fibroblasts. BJ-TERT cells were seeded at 50,000 cells/well and allowed to culture for 24 hours. Cells were then treated with and without Saponin. All cells were then stained with Calcein AM and EthD-1. Top: Cells without Saponin treatment. Bottom: Cells with Saponin treatment. Left: Calcein AM staining. Right: EthD-1 staining.

Detection

Size

Catalog Number

Colorimetric / Fluorometric

96 Assays

CBA-240

CytoSelect™ LDH Cytotoxicity Assay Loss of cell membrane integrity is one of the hallmarks of cytotoxicity. Upon cell death, lactate dehydrogenase (LDH) is released from the cytoplasm through the damaged membrane. Our CytoSelect™ LDH Cytotoxicity Assay provides a convenient plate-based method for testing cytotoxicity based on LDH release. In this assay, cells are cultured in a 96-well plate with and without the compound to be tested. LDH released into the media from cells converts a lactate substrate to pyruvate and generates NADH. In the presence of NADH, the colorimetric dye WST-1 is converted to a formazan that generates an orange color. Detection is performed in a standard colorimetric plate reader.

Product Name CytoSelect™ LDH Cytotoxicity Assay Kit

Phone 1-858-271-6500

LDH Release from HEK 293 Cells. 20,000 cells/well were cultured for 24 hours. After adding various concentrations of Triton X-100, the LDH Cytotoxicity Assay Reagent was added followed by a 30 minute incubation at 37ºC and 5% CO2. Detection

Size

Catalog Number

Colorimetric

960 Assays

CBA-241

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Cell Health

CELL-BASED ASSAYS

Cellular Senescence Assays Senescence Associated (SA) -galactosidase is a common biochemical marker of cellular senescence. Cells expressing such markers have been identified in vivo in tissues. SA ß-Gal catalyzes hydrolysis of X-gal, which produces a blue color in senescent cells. We offer three kit formats to test cellular senescence via SA-ß-galalactosidase activity:  Our ß-Gal Staining Kit allows you to visualize senescence by standard light microscope.  Our Quantitative Cellular Senescence Assay measures senescence in cells cultured in a 35mm dish by either flow cytometry or fluorescence microscopy  Our 96-Well Cellular Senescence Assay provides a higher throughput assay in a fluorescence plate reader. Recent Product Citation Malhotra, D. et al. (2010). Global mapping of binding sites for Nrf2 identifies novel targets in cell survival response through ChIP-Seq profiling and network analysis. Nucleic Acid Res. 10.1093/nar/gkq212. (CBA-231) Product Name

Detection

Cellular Senescence Assay Kit (SA -gal Staining)

Light Microscopy Fluorometric Plate Reader

96-Well Cellular Senescence Assay (SA -gal Activity)

Flow Cytometry / Fluorescence Microscopy

Quantitative Cellular Senescence Assay (SA -gal)

Size

Catalog Number

50 Assays

CBA-230

5 x 50 Assays

CBA-230-5

120 Assays

CBA-231

5 x 120 Assays

CBA-231-5

10 Assays

CBA-232

5 x 10 Assays

CBA-232-5

CytoSelect™ Anoikis Assays Anoikis is defined as death of adherent cells due to loss of adhesion to the extracellular matrix. Our Anoikis Assays allow you to quantify and monitor anchorage-dependent cell death using a precoated plate. Live cells can be viewed under a microscope and quantified on a plate reader by MTT (colorimetric) or Calcein AM (fluorometric), both included with the kit. Dead cells are detected with an EthD-1 reagent. Recent Product Citations 1. Lee, H.W. et al. (2013). Tpl2 kinase impacts tumor growth and metastasis of clear cell renal cell carcinoma. Mol. Cancer Res. 11:1375-1386. (CBA-080) 2. Sisto, M. et al. (2009). Fibulin-6 expression and anoikis in human salivary gland epithelial cells: implications in Sjogren's syndrome. Int. Immunol. 21:303-311. (CBA-080) 3. Liu, H. et al (2008). Cysteine-rich protein 61 and connective tissue growth factor induce de-adhesion and anoikis of retinal pericytes. Endocrinology 149:1666-1677. (CBA-080) Product Name

 Versatile: Detect live and dead cells by microscopy, fluorescence, or flow cytometry  Quantitative: Measure live and dead cells on a fluorescence plate reader; live cells may also be quantified on a standard microplate reader

Anoikis of Human Fibroblast BJ-TERT Cells. 50,000 cells/well were seeded in a control plate (left) and a Poly-HEMA coated plate (right) and cultured for 24 hours. Cells on the control plate were stained with Calcein AM. Cells on the Poly-HEMA coated plate were stained with EthD-1. Detection

Size

Catalog Number

CytoSelect™ 24-Well Anoikis Assay

Colorimetric / Fluorometric

24 Assays

CBA-080

CytoSelect™ 96-Well Anoikis Assay

Colorimetric / Fluorometric

96 Assays

CBA-081

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CELL-BASED ASSAYS

Cell Health

CytoSelect™ Fluorometric Cell Proliferation Assay Reagent Cell proliferation is easily measured by the addition of a variety of dyes that can be correlated with the number of cells. Various dyes producing a visible color are available to measure proliferation rates, but fluorometric dyes are often more sensitive and may be a superior choice for researchers with access to a fluorescence-based microplate reader. Our CytoSelect™ Cell Proliferation Assay Reagent (Fluorometric) provides a simple, single reagent method to measure proliferation of cells. The fluorometric dye is added directly to cultured cells. Upon entering metabolically active live cells, the nonfluorescent dye is converted to a bright red fluorescent form. Quantitation is performed using a fluorescence plate reader with excitation at 560 nm and emission at 590-600 nm. This reagent is versatile and can be used with a wide variety of cell types including cultured mammalian and piscine cells, bacteria, yeast, fungi, and protozoa.

Product Name CytoSelect™ Cell Proliferation Assay Reagent, Fluorometric

Human HEK 293 Cell Density. Cells were seeded at various densities in triplicate and allowed to culture for 24 hours. Cells were then treated with the CytoSelect™ Cell Proliferation Assay Reagent for 6 hours at 37ºC and 5% CO2.

Detection

Size

Catalog Number

Fluorometric

960 Assays

CBA-250

CytoSelect™ MTT Cell Proliferation Assay Cell proliferation is easily measured by the addition of a variety of dyes that produce a visible color that can be correlated with the number of cells. Our CytoSelect™ MTT Cell Proliferation Assay provides a simple method to measure proliferation of cells. The cell-permeable MTT dye is added directly to cultured cells followed by a detergent solution. Quantitation is performed using a standard microplate reader at 540-570 nm. Product Name CytoSelect™ MTT Cell Proliferation Assay

Detection

Size

Catalog Number

Colorimetric

960 Assays

CBA-252

CytoSelect™ WST-1 Cell Proliferation Assay Reagent Our CytoSelect™ WST-1 Cell Proliferation Assay provides a similar method to our MTT Cell Proliferation Assay, but with a single reagent format that does not require a detergent solubilization step. Quantitation is performed using a standard microplate reader at 450 nm. Product Name CytoSelect™ WST-1 Cell Proliferation Assay Reagent

Phone 1-858-271-6500

Detection

Size

Catalog Number

Colorimetric

960 Assays

CBA-253

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Cell Health

CELL-BASED ASSAYS

CytoSelect™ BrdU Cell Proliferation ELISA Kit BrdU is a thymidine analog that can incorporate into newly synthesized DNA strands of actively proliferating cells. Our CytoSelect™ BrdU Cell Proliferation ELISA Kit provides a convenient plate-based method to measure this incorporation. Once the BrdU is incorporated into the DNA, cells are fixed and DNA is denatured. Incorporated BrdU can be quantified in the denatured DNA by an anti-BrdU antibody. The absorbance readings at 450 nm can be directly correlated to cell proliferation.

Assay Principle for the CytoSelect™ BrdU Cell Proliferation ELISA Kit. Product Name CytoSelect™ BrdU Cell Proliferation ELISA Kit

Detection

Size

Catalog Number

Colorimetric

96 Assays

CBA-251

CytoSelect™ Proliferating Cell Nuclear Antigen (PCNA) ELISA Kit Proliferating Cell Nuclear Antigen (PCNA) acts as a processivity factor for DNA polymerase by associating with various proteins involved in DNA replication. It is also associated with chromatin remodeling and cell cycle control, and it is often used as a marker of cell proliferation. Our CytoSelect™ PCNA ELISA Kit provides a convenient plate-based method to quantify PCNA levels in nuclear or whole cell extracts.  Sensitive: Detect PCNA as low as 12.5 ng/mL  Versatile: Measure PCNA levels from human, mouse, or rat whole cell lysates or nuclear extracts  Quantitative: Measure results in a colorimetric plate reader against a provided PCNA standard

Product Name CytoSelect™ Proliferating Cell Nuclear Antigen (PCNA) ELISA Kit

PCNA Detection in HeLa Whole Cell Lysates. Whole cell lysates were prepared in a RIPA lysis buffer. Protein concentrations were determined by BCA protein assay. Detection

Size

Catalog Number

Colorimetric

96 Assays

CBA-254

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CELL-BASED ASSAYS

Cell Hypoxia

HIF-1 Alpha DNA Binding Activity Assay Kit Cell hypoxia, or low oxygen condition, is a normal physiological response to certain body stressors such as high altitudes, but it can also be a symptom of pathological conditions and is often used as a marker for tumor cells. In response to hypoxic conditions, the hypoxia-inducible factor 1 transcriptional activator complex (HIF-1) plays a role in activating several hypoxia-responsive genes such as erythropoietin and VEGF. During hypoxia, the alpha subunit of HIF-1 accumulates and translocates from the cytosol to the nucleus, where it dimerizes with the beta subunit and becomes transcriptionally active. It then binds transcriptional coactivators to induce gene expression. The HIF-1 Alpha DNA Binding Activity Assay Kit detects activated HIF-1 in an ELISA format. Active HIF1 complex is captured on a double-stranded oligo containing a hypoxic response element (HRE) that is attached to the plate. Detection is then performed with a primary antibody followed by an HRPconjugated secondary antibody. The assay will detect HIF-1 complexes from human, mouse or rat protein samples. Product Name HIF-1 Alpha DNA Binding Activity Assay Kit

Detection Specificity of HIF-1 Alpha. HeLa cells were incubated in the presence or absence of 0.2 mM deferoxamine mesylate (DFO) for 4 hours at 37ºC. Nuclear extracts were prepared using the Nuclear/Cytosolic Fractionation Kit (#AKR-171). 100 pmol of non-biotinylated wild type or mutated HRE double stranded competitor oligos were added to the Complete DNA Binding Buffer just prior to inclusion in the assay. Detection

Size

Catalog Number

Colorimetric

96 Assays

CBA-282

HIF-1 Alpha ELISA Kits Our HIF-1 Alpha ELISA Kits provide a convenient method for detection and quantitation of human, mouse, or rat HIF-1 Alpha in cells or tissues. Two ELISA kit formats are available:  The HIF-1 Alpha Sandwich ELISA Kit detects HIF -1 Alpha in any protein sample including tissue homogenates, whole cell lysates, or nuclear extracts. Samples are added to an anti-HIF-1 Alpha antibody coated plate. Quantitation of unknown samples is performed by comparison of the OD values to those of a known standard.  The HIF-1 Alpha Cell Based ELISA Kit allows the detection of HIF-1 Alpha levels in intact cells. Cells are seeded in a tissue culture treated plate suitable for reading in a 96-well plate-based luminometer. Cells are fixed and permeabilized to allow detection with the anti-HIF-1 antibody. Detection is performed by chemiluminescence. Product Name

Detection of Nuclear HIF-1 Alpha with the HIF-1 Alpha Sandwich ELISA Kit. HeLa cells were incubated in the presence or absence of 0.2 mM DFO for 4 hours at 37ºC. HIF-1 Alpha levels were measured in untreated (blue bars) and treated (red bars) nuclear extracts according to the Assay Protocol. Detection

Size

Catalog Number

HIF-1 Alpha Sandwich ELISA Kit

Colorimetric

96 Assays

CBA-280

HIF-1 Alpha Cell Based ELISA Kit

Chemiluminescent

96 Assays

CBA-281

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Adipogenesis

CELL-BASED ASSAYS

CytoSelect™ 96-well Adipogenesis Assay Kit The ability to regulate the cell cycle and differentiation of adipocytes is important to the understanding of obesity. Adipogenesis is the process in which preadipocytes develop into mature adipocytes in a multistep process that requires the sequential activation of numerous transcription factors. The 3T3-L1 cell line is the best characterized model for adipogenesis in vitro. 3T3-L1 cells display a fibroblast-like phenotype when grown under normal conditions. However, when treated with a combination of IBMX, insulin, and dexamethasone, these cells undergo terminal differentiation resulting in a more rounded phenotype and the formation of intracellular lipid droplets. The CytoSelect™ 96-Well Adipogenesis Assay quantitatively measures lipid droplet accumulation in cultured cells of the 3T3-L1 model. Quantitation is performed either in a standard colorimetric plate reader with Oil Red O stain, or in a fluorescence plate reader with Nile Red fluorometric stain.

Staining of 3T3-L1 Cells with Oil Red O. 20,000 cells/well of preadipocyte 3T3-L1 cells were seeded overnight in a 96-well plate. Cells were uninduced (top) or induced (bottom) for 7 days and stained with Oil Red O colorimetric stain according to the Assay Protocol. Product Name CytoSelect™ 96-Well Adipogenesis Assay Kit

Staining of 3T3-L1 Cells with Nile Red Fluorescent Stain. 20,000 cells/well of preadipocyte 3T3-L1 cells were seeded overnight in a 96-well plate. Cells were uninduced (top) or induced (bottom) for 7 days and stained with Nile Red Fluorescent Stain according to the Assay Protocol. Detection

Size

Catalog Number

Colorimetric / Fluorometric

200 Assays

CBA-290

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info@cellbiolabs.com

CELL-BASED ASSAYS

Phagocytosis

CytoSelect™ 96-Well Phagocytosis Assays Phagocytosis may be assayed by measuring the engulfing of a cell “substrate” such as an erythrocyte (RBC) or Zymosan particle. Traditional phagocytosis assays involve manually counting the engulfed substrates under a microscope. This process is tedious and time-consuming, can be somewhat inaccurate, and is not amenable to high throughput. CytoSelect™ 96-Well Phagocytosis Assays are more accurate, high-throughput alternatives to the standard phagocytosis assay. The assays may be adapted for use in 48-well and 24-well plates if desired.

Assay Principle for the CytoSelect™ 96-Well Phagocytosis Assay (Zymosan). Product Name

 Highly Accurate: Eliminates manual counting  High Throughput: 96-well plate format  Quantitative: Measure OD in a standard microplate reader  Flexible: Choose from 3 substrates: E. coli, Zymosan particles, or red blood cells* *Red blood cells are not provided in the kit. Fresh RBCs should be obtained immediately prior to running the assay. E. coli and Zymosan particles are provided in their respective kits. Recent Product Citations 1. Lee, J.K. et al. (2010). Regulator of G-protein signaling-10 negatively regulates NF-kB in microglia and neuroprotects dopaminergic neurons in hemiparkinsonian rats. J. Neurosci. 31:11879-11888. (CBA-220) 2. Winnicka, B. et al. (2010). CD13 is dispensible for normal hematopoiesis and myeloid cell functions in the mouse. J. Leukoc. Biol. 88(2):347-359. (CBA-220) 3. Hamilton, C.M. et al. (2009). Fasciola hepatica tegumental antigen suppresses dendritic cell maturation and function. Infect. Immun. 77:2488-2498. (CBA-220) 4. Dowling, D.J. et al. (2009). Major secretory antigens of the helminth Fasciola hepatica activate a suppressive dendritic cell phenotype that attenuates Th17 cells but fails to activate Th2 immune responses. Infect. Immun. 78:793-801. (CBA-220) 5. Pierce, L.M. et al. (2012). Effect of heavy metal tungsten alloy particles on oxidative product formation and phagocytosis by lung macrophages. Am. J. Respir. Crit. Care Med. 185:A4666. (CBA-224) 6. Polancec, D.S. et al. (2012). Azithromycin drives in vitro GMCSF/IL-4-induced differentiation of human blood monocytes toward dendritic-like cells with regulatory properties. J. Leukoc. Biol. 91:229.-243. (CBA-224)

Particle Engulfment with the CytoSelect™ 96-Well Phagocytosis Assay (Zymosan). Detection

Size

Catalog Number

CytoSelect™ 96-Well Phagocytosis Assay (E. coli)

Colorimetric

96 Assays

CBA-222

CytoSelect™ 96-Well Phagocytosis Assay (Red Blood Cell)

Colorimetric

96 Assays

CBA-220

96 Assays

CBA-224

CytoSelect™ 96-Well Phagocytosis Assay (Zymosan)

Colorimetric

5 x 96 Assays

CBA-224-5

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Cell Contraction and Angiogenesis

CELL-BASED ASSAYS

Collagen-Based Cell Contraction Assay Wound healing is comprised of epithelialization, connective tissue deposition, and contraction. The contraction process is believed to be mediated by specialized fibroblasts (myofibroblasts). 3D collagen gels have been widely used in fibroblast contraction studies. Our Cell Contraction Assay provides a simple system to assess cell contractivity and to screen for cell contraction mediators. The system uses a 3D collagen matrix to measure changes in the collagen gel size. An optional contraction inhibitor is provided. Recent Product Citations 1. Kotio, K.U. et al. (2011). Implication of microRNAs in atrial natriuretic peptide and nitric oxide signaling in vascular smooth muscle cells. Am. J. Physiol. Gastrointest. Liver Physiol. 301: C929C937. 2. Schell, C. et al. (2010). 15-deoxy-delta12-14-prostaglandin-J2 induces hypertrophy and loss of contractility in human testicular peritubular cells: implications for human male fertility. Endocrinology 151:12571268.

Product Name Cell Contraction Assay

Assay Principle for the Collagen-Based Cell Contraction Assay.

Detection

Size

Catalog Number

Light Microscopy

24 Assays

CBA-201

Endothelial Tube Formation (In Vitro Angiogenesis) Assay For angiogenesis to occur, endothelial cells must escape their stable location and break through the basement membrane. These cells proliferate to form new blood vessels. Our Endothelial Tube Formation Assay provides an easy, robust system to assess angiogenesis in vitro. The assay uses an ECM gel matrix derived from mouse sarcoma cells; this matrix very closely resembles an in vivo basement membrane environment. Recent Product Citations 1. Yu, J.G. et al. (2013). Baroreflex deficiency hampers angiogenesis after myocardial infarction via acetylcholine-alpha7-nicotinic ACh receptor in rats. Eur. Heart J. 34:2412-2420. 2. Bid, H. et al. (2013). Dual targeting of the Type 1 insulin-like growth factor receptor and its ligands as an effective antiangiogenic strategy. Clin. Cancer Res. 19:2984-2994. 3. Cai, C. et al. (2012). SIVmac239-Nef downregulates cell surface expression of CXCR4 in tumor cells and inhibits proliferation, migration and angiogenesis. Anticancer Res. 23:2759-2768. 4. Bid, H. et al. (2012). Potent inhibition of angiogenesis by the IGF -1 receptor-targeting antibody SCH717454 is reversed by IGF-2. Mol. Cancer Ther. 11:649-659. Product Name Endothelial Tube Formation Assay (In Vitro Angiogenesis)

HUVEC Tube Formation on ECM Gel. HUVEC cells from a standard tissue culture plate were incubated on an ECM gel. After several hours tube formation can be visualized under a light microscope. Detection

Size

Catalog Number

Light Microscopy

50 Assays

CBA-200

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CELL-BASED ASSAYS

Autophagy

GFP-LC3 Expression Vectors MAP LC3 is the most published autophagosome marker protein. LC3 associates to the inner and outer limiting membranes of the autophagosome. There are two forms of LC3 visible by immunoblot: LC3I which is found in the soluble fraction, and LC3II which is found in the membrane fraction. The proportion of LC3II increases during autophagy. Our GFP-LC3 expression vectors are convenient tools for the study of autophagy. These vectors are available in three formats: mammalian, lentiviral, and retroviral expression vectors. Each vector contains a GFP reporter gene. In addition, a GFP control plasmid is provided at no additional charge.

Recent Product Citations 1. Chen, W. et al. (2012). Andrographolide induces autophagic cell death in human liver cancer cells through cyclophilin D-mediated mitochondrial permeability transition pore. Carcinogenesis 10.1093/carcin/bgs264. (CBA-401) 2. Cina, D.P. et al. (2012). Inhibition of MTOR disrupts autophagic flux in podocytes. J. Am. Soc. Nephrol. 23:412-420. (CBA-401) 3. Tu, S.P. et al. (2011). IFN-gamma inhibits gastric carcinogenesis by inducing epithelial cell autophagy and T-cell apoptosis. Cancer Res. 71:4247-4259. (CBA-401) Product Name

Size

Catalog Number

pCMV-GFP-LC3 Expression Vector

100 µL

CBA-401

pSMPUW-GFP-LC3 Lentiviral Expression Vector

10 µg

LTV-801

pMXs-GFP-LC3 Retroviral Expression Vector

10 µg

RTV-801

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

STEM CELL RESEARCH

Induced Pluripotent Stem Cells

iPS Cell Reprogramming Reprogramming of adult cells into induced pluripotent stem cells (iPS) has provided an important new vehicle to facilitate stem cell research. Recent studies have shown that this may be accomplished by the introduction of key genes into somatic cells by transduction with various viral vectors or transfection of plasmids. Retroviral and lentiviral vectors appear to achieve among the highest levels of efficiency of iPS cell generation. We offer an extensive collection of vectors for iPS cell reprogramming.

Retroviral Vectors and Packaging Cells for iPS Cell Generation Our iPS retroviral vectors are constructed from the pMXs vector backbone developed by Dr. Toshio Kitamura at the University of Tokyo.* Each vector contains one of 6 factors shown to help reprogram adult fibroblasts into iPS cells. Both human and mouse genes are available individually or in sets. Separate retroviral vectors are available for p53 shRNA, which has been shown to potentially increase the efficiency of iPS cell generation. Platinum Retroviral Packaging Cells provide an easy way to produce high-titer retroviruses from these stem cell plasmids. For additional information on these cell lines please see p. 64. *Kitamura, T. et al. (2003). Exp. Hematol. 31:1007-1014.

Human iPS Vectors

Mouse iPS Vectors Vector Backbone

Catalog Number

Oct-3/4

pMXs

RTV-705

RTV-702

Sox2

pMXs

RTV-706

pMXs

RTV-703

c-Myc

pMXs

RTV-707

Klf4

pMXs

RTV-704

Klf4

pMXs

RTV-708

NANOG

pMXs

RTV-709

NANOG

pMXs

RTV-711

Lin28

pMXs

RTV-710

Lin28

pMXs

RTV-712

Set of 4 vectors (Oct-3-4, Sox2, c-Myc, Klf4)

pMXs

RTV-701-C

Set of 4 vectors (Oct-3-4, Sox2, c-Myc, Klf4)

pMXs

RTV-705-C

Set of 6 vectors (Oct-3-4, Sox2, c-Myc, Klf4, NANOG, Lin28)

pMXs

RTV-709-C

Set of 6 vectors (Oct-3-4, Sox2, c-Myc, Klf4, NANOG, Lin28)

pMXs

RTV-711-C

p53 shRNA

pRetro

RTV-410

p53 shRNA

pRetro

RTV-400

Target Name

Vector Backbone

Catalog Number

Oct-3/4

pMXs

RTV-701

Sox2

pMXs

c-Myc

Target Name

Retroviral Packaging Cell Lines Product Name

Size

Platinum-E Retroviral Packaging Cell Line, Ecotropic

>3 x 10 cells

RV-101

Platinum-A Retroviral Packaging Cell Line, Amphotropic

>3 x 106 cells

RV-102

Platinum-GP Retroviral Packaging Cell Line, Pantropic pCMV-VSV-G Packaging Vector (for use with Platinum-GP cells)

Catalog Number

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

>3 x 10 cells

RV-103

10 Âľg

RV-110

Fax 1-858-271-6514

Induced Pluripotent Stem Cells

STEM CELL RESEARCH

Polycistronic Vectors for iPS Cell Generation Our Polycistronic Viral Vectors provide a convenient way to generate iPS cells. The defined stem cells factors Klf4, Oct-3/4, Sox2 and c-Myc are in-frame fused into a single open reading frame (ORF) by selfcleaving 2A peptides. The transcription factor ORF is followed by IRES-GFP as a reporter to verify viral transduction into your target cell. Efficiencies of iPS generation are typically higher compared to transduction of four separate viruses each containing a single gene.

 More Efficient: Up to 10-fold higher efficiency compared to multi-virus transduction, and 500-fold compared to non-viral methods  Reporter Convenience: GFP reporter gene helps to monitor viral transduction

Two vectors are available:  pLentG-KOSM is a lentiviral vector containing mouse sequences  pRetroG-OKSM is a retroviral vector containing human sequences

Expression of Stem Cell Factors and GFP. Top: Transient expression of KOSM fusion gene in 293T cells confirmed by Western blot. Bottom: GFP fluorescence in MEF cells 3 days after infection with lentivirus containing KOSM fusion.

Open Reading Frame of pLentG-KOSM Lentiviral Vector.

Characterization of iPS Cell Colonies Generated from MEFs Infected with Lentivirus Containing the KOSM Fusion. Top: Staining of pluripotency markers in induced cell colonies at 200x magnification. Bottom: AP staining at 100x magnification and morphology at 40x magnification in induced cell colonies.

Product Name

Size

Catalog Number

pLentG-KOSM Polycistronic Lentiviral Vector (Mouse genes)

100 µL

LTV-700

pRetroG-OKSM Polycistronic Retroviral Vector (Human genes)

100 µL

RTV-700

For efficient packaging of your virus, please see our Lentiviral Packaging Systems on p. 58 and Retroviral Packaging Cell Lines on p. 64 and 67.

www.cellbiolabs.com

info@cellbiolabs.com

STEM CELL RESEARCH

Retroviral Expression Systems

Platinum Retroviral Expression Systems for Stem Cells Retroviral vectors are useful for delivering genes of interest into a host cell where integration into the genome is desired. However, traditional retroviral expression technologies usually result in low viral titers which make gene expression studies difficult. Our Platinum Retroviral Expression Systems incorporate superior packaging cell lines and vector technologies to produce high-titer virus with a single plasmid transfection. The Platinum Expression Systems include one of our exclusive Platinum Packaging Cell Lines which already contain the gag and pol genes; the Ecotropic and Amphotropic cells also contain an envelope protein. Simply clone your gene of interest into the vector provided and transfect into the Platinum cells. If you choose a Pantropic system, simply co-transfect with the VSV-G plasmid provided. The Platinum Expression Systems below are specially designed for superior expression with either ES/ EC cells or hematopoietic stem cells. For more information on our Platinum Expression Systems for a variety of cells, please see page 60.

Ecotropic

Pantropic

Human

+++

N.S.

+++

Mouse

+++

Rat

+++

Monkey

+++

N.S.

+++

Cat

+++

N.S.

+++

Dog

+++

N.S.

+++

N.S.

+++

Bird

N.S.

+++

Fish

N.S.

+++

Frog

N.S.

+++

Insect

N.S.

+++

Mollusk

N.S.

+++

Hamster

*Virus must be packaged with a pantropic envelope protein such as VSVG.

 Higher Viral Yields: Average titer 107 infectious units/mL with transient transfection  Versatile: 3 Packaging cell lines for use with nearly any target host species  Optimized for Stem Cell Studies: Specially designed expression systems for either ES/EC cells or hematopoietic stem cells Catalog Number

Amphotropic

N.S. = Not Suitable Suitability of Platinum Retroviral Expression Systems by Host Species.

Packaging Cell Line

Transfer Vector

Envelope Vector

Control Vector

VPK-303

Plat-E (Ecotropic)

pMCs-Puro

———

pMCs-GFP

VPK-304

Plat-A (Amphotropic)

pMCs-Puro

———

pMCs-GFP

VPK-305

Plat-GP (Pantropic)

pMCs-Puro

pCMV-VSV-G

pMCs-GFP

VPK-306

Plat-E (Ecotropic)

pMYs-Puro

———

pMYs-GFP

VPK-307

Plat-A (Amphotropic)

pMYs-Puro

———

pMYs-GFP

VPK-308

Plat-GP (Pantropic)

pMYs-Puro

pCMV-VSV-G

pMYs-GFP

Components of the Platinum Retroviral Expression Systems for Stem Cells.

Product Name

Size

Catalog Number

Platinum ES/EC Retroviral Expression System, Ecotropic

1 kit

VPK-303

Platinum ES/EC Retroviral Expression System, Amphotropic

1 kit

VPK-304

Platinum ES/EC Retroviral Expression System, Pantropic

1 kit

VPK-305

Platinum HSC Retroviral Expression System, Ecotropic

1 kit

VPK-306

Platinum HSC Retroviral Expression System, Amphotropic

1 kit

VPK-307

Platinum HSC Retroviral Expression System, Pantropic

1 kit

VPK-308

Phone 1-858-271-6500

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Feeder Cells

STEM CELL RESEARCH

Stem Cell Feeders Leukemia inhibitory factor (LIF) is useful for maintaining the undifferentiated state of mouse embryonic stem (mES) cells. However, LIF does not have the same effect on human embryonic stem (hES) cells. Therefore, hES cells require the use of feeder cells for both derivation and maintenance. We offer a variety of feeder cells for stem cell culture. All feeder cells must be mitotically inactivated prior to use.

SNL 76/7 Passage-Independent Feeder Cells for iPS Culture The SNL 76/7 is an immortalized cell line derived from mouse fibroblast STO cells which have been transformed with murine LIF and neomycin resistance genes.  Superior Culture: Transformed with LIF gene for better maintenance of undifferentiated state  Versatile: Useful for culture of human and mouse iPS cells and as a feeder for ES cells  Passage-Independent: Immortalized cell line

Recent Product Citations 1. Osakada, F. et al (2009). In vitro differentiation of retinal cells from human pluripotent stem cells by small-molecule induction. J. Cell Sci. 132:3169-3179. 2. Liu, Y. et al. (2009). Zeb1 represses Mitf and regulates pigment synthesis, cell proliferation, and epithelial morphology. Invest. Ophthalmol. Vis. Sci. 50:5080-5088. 3. Tsubooka, N. et al. (2009). Roles of Sall4 in the generation of pluripotent stem cells from blastocytes and fibroblasts. Genes Cells 14:683-694. 4. Lan, Z-J. et al. (2009). Extra-germ cell expression of mouse nuclear receptor subfamily 6, group A, member 1 (NR6A1). Biol. Reprod. 80:905-912.

Product Name

Size 6

3 x 10 cells

SNL Feeder Cells

Catalog Number CBA-316

JK1 Passage-Independent Feeder Cells JK1 is an immortalized CD34+ stromal cell line that supports long-term proliferation of stem cells. It has been shown to maintain capacity for stem cell renewal even after serial passaging for over one year. JK1 may be used for culture of a variety of cell types including pluripotent ES cells, germ-line derived stem cells such as SPCs and MASCs, and primordial germ cell-derived EG cells. Product Name

JK1 Cells Support Maintenance of mES Cells. Immunofluorescence staining of germ cell nuclear antigen (GCNA). Antibody staining is green; nuclear counterstain is blue.

Size 6

1 x 10 cells

JK1 Feeder Cells

Catalog Number CBA-315

MEF Feeder Cells Our murine embryonic fibroblast (MEF) feeder cells are useful for the maintenance of human or mouse ES cells in their undifferentiated state. Cells must be mitotically inactivated prior to use. Product Name

Size

CBA-310

CBA-313

5 x 10 cells

MEF Feeder Cells

5 x 10 cells

MEF Feeder Cells, Hygromycin-resistant

Catalog Number

MEF Feeder Cells, Neomycin-resistant

5 x 10 cells

CBA-311

MEF Feeder Cells, Puromycin-resistant

5 x 106 cells

CBA-312

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STEM CELL RESEARCH

Colony Formation Assays

CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay Hematopoietic stem cells (HSCs), when cultured in a suitable semisolid matrix such as methylcellulose supplemented with cytokines & nutrients, proliferate to form discrete cell clusters or colonies. Such HSCs or hematopoietic progenitors are known as colonyforming cells (CFCs). In classic CFC assays, cells are cultured in a 35mm dish for 14-21 days so the colonies can reach a certain size for manual counting, which can be tedious and subjective. The CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay provides a high-throughput method to quantify CFCs in just 7-10 days with no manual cell counting required. Cells are lysed, solubilized, and quantified using a fluorescent dye included in the kit. Alternatively, cells may be recovered for further culture and analysis.  Fast Results: 7-10 days vs. 2-3 weeks  Plate Reader Convenience: Eliminates manual counting  Easier Reagent Handling: Methylcellulose media can be handled using a pipet instead of a syringe Assay Principle for the CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay.

HSC Colony Formation. Human bone marrow derived CD34+ Hematopoietic Progenitor Cells were seeded at 3000 cells/well and cultured for 7-10 days in the absence or presence of growth factors/cytokines. Colonies were quantified according to the assay protocol. A: After 7 days without cytokines. B: After 7 days in presence of cytokines. C: After 10 days in presence of cytokines (hemoglobin visible). Product Name

Detection

CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay

Phone 1-858-271-6500

Fluorometric

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

96 Assays

CBA-320

5 x 96 Assays

CBA-320-5

Fax 1-858-271-6514

Colony Formation Assays

STEM CELL RESEARCH

StemTAG™ 96-Well Stem Cell Colony Formation Assay Our StemTAG™ 96-Well Stem Cell Colony Formation Assay provides a high-throughput method to quantify ES cells in just 7-10 days with no manual cell counting required. After colonies are formed, stem cells may be analyzed in 3 ways:

 Fast Results: 7-10 days vs. 2-3 weeks using conventional methods  Versatile: Quantify cells using fluorescent dye, measure alkaline phosphatase activity, or recover cells for further analysis  Plate Reader Convenience: No manual cell counting required

1. Lyse cells, then quantify using a fluorescent dye included in the kit. 2. Lyse cells, then measure alkaline phosphate activity using reagents provided. 3. Recover colonies for further culture and analysis. This assay may be of particular interest for the study of tumor stem cells.

StemTAG™ Stem Cell Colony Formation Assay Principle. Product Name

Anchorage-Independent Growth of Mouse ES-D3 Cells. Top: Phase Contrast. Bottom: Alkaline Phosphatase Staining. Detection

StemTAG™ 96-Well Stem Cell Colony Formation Assay

Fluorometric

www.cellbiolabs.com

Size

Catalog Number

96 Assays

CBA-325

5 x 96 Assays

CBA-325-5

info@cellbiolabs.com

STEM CELL RESEARCH

Alk Phos Assays, Primers, RNA/Protein

StemTAG™ Alkaline Phosphatase Kits The StemTAG™ Alkaline Phosphatase Staining and Activity Assay Kits monitor AP activity via both immunocytochemistry staining and a colorimetric 96-well plate-based activity assay. The staining and activity assay kits are also sold separately.

StemTAG™ Alkaline Phosphatase Staining Kit. Murine embryonic stem cells (ES-D3) were maintained in an undifferentiated state with LIF. To induce differentiation, LIF was withdrawn over several days. Various differentiation events were observed: cells became flattened and enlarged with reduced proliferation. On day 5, cells were stained according to the assay protocol. Product Name

 Fast Results: Staining and Activity Assay protocols each take less than 1 hour  Versatile: Useful for human ES, EG and EC cells, as well as mouse ES and EG cells

Recent Product Citations 1. Lee, J. et al. (2010). Ultraviolet A regulates adipogenic differentiation of human adipose tissue-derived mesenchymal stem cells via up-regulation of kruppel-like factor 2. J. Biol. Chem. 285:32647-32656. (CBA-300) 2. Izadyar, F. et al. (2008). Generation of multipotent cell lines from a distinct population of male germ line stem cells. Reproduction 135:771-784. (CBA-300) 3. Kim, Y. et al. (2009). Cyclin-dependent kinase 2-associating protein 1 commits murine embryonic stem cell differentiation through retinoblastoma protein regulation. J. Biol. Chem. 284:23405-23414. (CBA-301) 4. Yue, Y. et al. (2008). A single intravenous injection of adenoassociated virus serotype-9 leads to whole body skeletal muscle transduction in dogs. Mol. Ther. 16(12):1944-1952. (CBA-301) 5. Ghosh, A. et al (2007). Efficient whole-body transduction with trans-splicing AAV vectors. Mol. Ther. 15(4):750-755. (CBA-301) Detection

Size

Catalog Number

ICC & Colorimetric

2 x 100 Assays

CBA-302

ICC & Fluorometric

2 x 100 Assays

CBA-308

Colorimetric

100 Assays

CBA-301

Fluorometric

100 Assays

CBA-307

StemTAG™ Alkaline Phosphatase Staining Kit (Red)

ICC

100 Assays

CBA-300

StemTAG™ Alkaline Phosphatase Staining Kit (Purple)

ICC

100 Assays

CBA-306

StemTAG™ Alkaline Phosphatase Staining and Activity Assay Kit

StemTAG™ Alkaline Phosphatase Activity Assay Kit

StemTAG™ PCR Primer Set for Stem Cell Characterization Pluripotent stem cells can differentiate into cells derived from all three embryonic germ layers: endoderm, mesoderm and ectoderm. Our StemTAG™ PCR Primer Set provides an efficient system for monitoring ES cell differentiation/undifferentiation. Seven primer sets are included: primers for two widely studied stem cell markers (Oct-4 and NANOG), one marker for each embryonic germ layer (AFP/Endoderm, Flk-1/Mesoderm and NCAM/Ectoderm), and two controls (GAPDH and ß-Actin). Primers are suitable for either end-point or real-time (quantitative) PCR. Product Name StemTAG™ PCR Primer Set for Stem Cell Characterization

Size

Catalog Number

50 Reactions

CBA-303

Total RNA and Protein from Murine ES-D3 Cell Line

Product Name

Size

Catalog Number

Total RNA—Murine Embryonic Stem Cell Line D3

50 µg

CBA-304

Total Protein—Murine Embryonic Stem Cell Line D3

500 µg

CBA-305

Phone 1-858-271-6500

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VIRAL EXPRESSION

Viral Vector Overview

Recombinant Viral Gene Delivery Recombinant viral vectors provide a powerful means of delivering a gene into a target cell. There are many viral vectors available, and there are pros and cons to each. Use the following table to select the best viral vector for your research.

Comparison of Viral Vectors for Gene Delivery Adeno-Associated Virus (AAV) (p. 41-48)

Adenovirus (p. 49-56)

Lentivirus (HIV-1, FIV, SIV) (p. 57-63)

Retrovirus (MMLV) (p. 64-72)

Transient or Stable

Transient

Transient or Stable

Stable

Will Infect Dividing Cells

Yes

Will Infect Non-Dividing Cells

Yes

Integrates into Target Cell Genome

No*

Yes

Immune Response in Target Cells

Very Low

High

Low

Moderate

Relative Viral Titer

XXX

XXXX

XXX

Relative Transduction Efficiency

XXX

XXXX

XXX

Gene Expression

*Native AAV can integrate, but recombinant AAV rarely does.

Typical Workflow for Viral Gene Delivery

Clone Gene of Interest

Viral Expression Plasmid

Package Virus

Packaging Plasmids and Cells

Measure Titer

Viral Quantitation Kit

Concentrate & Purify

Purification & Concentration Kit

Infect Target Cell

Viral Transduction Reagents

Cell Biolabs offers kits and reagents for every step in your workflow.

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

AAV Expression

VIRAL EXPRESSION

Adeno-Associated Virus Kits & Reagents Adeno-associated virus (AAV) is less immunogenic than adenovirus or retrovirus. We offer a comprehensive line of AAV kits and reagents to ensure you get the best expression from your AAV expression studies:  Helper Free Expression Systems  Premade AAV Controls  Helper Free Packaging Systems  Purification Kits  Expression & Control Vectors  Quantitation / Titer Kit  Viral Packaging Cell Line  Transduction Kits

AAV Helper Free Systems Production of recombinant AAV requires certain genes from the adenovirus genome, which means that an adenovirus usually needs to be present. The AAV Helper Free System eliminates the need for a helper adenovirus. Most of the required adenoviral genes (E2A, E4 and VA RNA) are provided in a pHelper plasmid, while the required E1 gene is provided by the 293 packaging cells.

 Safer: pHelper plasmid eliminates the need for a helper virus  Flexible: Packaging vectors and expression vectors available separately or as one complete system, so you only order what you need  Expandable: All plasmids are provided individually, not in a mixture, so you can amplify in competent cells

AAV Helper Free Systems are available for a variety of formats and serotypes:  AAV Complete Expression Systems

contain all packaging plasmids plus an expression vector and a GFP control vector: p. 42-45  AAV Packaging Systems contain the pHelper plasmid and a serotype-specific Rep-Cap plasmid for use with your own expression construct: p. 45  If you have an AAV packaging system for one serotype and want to try another, choose one of 8 different AAV Rep-Cap Plasmids from native serotypes 1 through 6 plus AAV-DJ and AAV-DJ/8: p. 45  If you already have an AAV packaging system and need a cloning vector, choose one of 10 different AAV Expression Vectors available individually: p. 46  Want to make a control virus? Choose one of our AAV Control Vectors: p. 46

Gene Delivery using the AAV Helper Free System.

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VIRAL EXPRESSION

AAV Expression

AAV Helper Free Complete Expression Systems AAV Helper Free Complete Expression Systems contain everything you need to produce high-titer recombinant adeno-associated virus:  pHelper Plasmid  Rep-Cap Plasmid (serotype specific)  GFP Control Vector  Choice of 10 AAV Expression Vectors:  Gene Expression (CMV or no promoter)  shRNA (U6 promoter)  Self complementary (scAAV)

AAV Helper Free Expression Systems are available for the following serotypes:  Native serotypes 1-6  AAV-DJ, engineered by DNA family shuffling to form a hybrid capsid from 8 different native serotypes; provides significantly higher infectivity rates in vitro (see table below)  AAV-DJ/8, a mutant of AAV-DJ that exhibits increased uptake in brain and other tissues in vivo, similar to serotypes 8 and 9

Cell Line

Cell or Tissue Source

AAV-1 AAV-2 AAV-3 AAV-4 AAV-5 AAV-6

AAV-8 AAV-9

AAVDJ

AAVDJ/8

Huh-7

Hu Liver

100

2.5

0.0

0.1

0.7

0.0

500

0.2

HEK293

Hu Kidney

100

2.5

0.1

0.7

0.1

500

0.3

HeLa

Hu Cervix

100

2.0

0.1

3.7

HepG2

Hu Liver

100

16.7

0.3

1.7

1.0

0.2

0.1

667

0.2

0.3

1250

0.5

Hep1A

Ms Liver

100

0.2

1.0

0.1

1.0

0.2

0.0

400

0.1

911

Hu Retina

100

11.1

0.2

0.1

500

0.0

CHO

Hm Ovary

100

14.3

1.4

333

10.0

1.0

25000

5.0

COS

Si Kidney

MeWo

Hu Skin

100

3.3

5.0

2.0

0.5

500

0.3

100

0.3

6.7

1.0

0.2

2857

1.0

NIH3T3

Ms Fibroblasts

100

2.9

0.3

500

0.1

A549

Hu Lung

100

0.5

0.1

1000

0.1

HT1180

Hu Fibroblasts

100

10.0

0.1

0.3

0.5

0.1

333

0.2

Monocytes

Hu Primary Monocytes

1111

100

125

1429

100

Immature DC

Hu Monocyte-derived DC

2500

100

222

2857

200

Mature DC

Hu Monocyte-derived DC

2222

100

333

3333

100

Relative Infectivity Rates of AAV Serotypes. Normalized to AAV-2 = 100. ND = Not determined.

AAV-DJ Helper Free Complete Expression Systems

Product Name

Size

Catalog Number

AAV-DJ Helper Free Expression System

1 kit

VPK-410-DJ

AAV-DJ Helper Free Bicistronic Expression System (Puro)

1 kit

VPK-415-DJ

AAV-DJ Helper Free Bicistronic Expression System (Neo)

1 kit

VPK-416-DJ

AAV-DJ Helper Free Bicistronic Expression System (Hygro)

1 kit

VPK-417-DJ

AAV-DJ Helper Free Bicistronic Expression System (GFP)

1 kit

VPK-418-DJ

AAV-DJ Helper Free Bicistronic Expression System (Blasticidin)

1 kit

VPK-419-DJ

AAV-DJ Helper Free Promoterless Expression System

1 kit

VPK-411-DJ

AAV-DJ Helper Free shRNA Expression System (Puro)

1 kit

VPK-412-DJ

AAV-DJ/Helper Free shRNA Expression System (GFP)

1 kit

VPK-413-DJ

scAAV-DJ Helper Free Expression System

1 kit

VPK-430-DJ

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AAV Expression

VIRAL EXPRESSION

AAV-DJ/8 Helper Free Complete Expression Systems Product Name

Size

Catalog Number

AAV-DJ/8 Helper Free Expression System

1 kit

VPK-410-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (Puro)

1 kit

VPK-415-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (Neo)

1 kit

VPK-416-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (Hygro)

1 kit

VPK-417-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (GFP)

1 kit

VPK-418-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (Blasticidin)

1 kit

VPK-419-DJ-8

AAV-DJ/8 Helper Free Promoterless Expression System

1 kit

VPK-411-DJ-8

AAV-DJ/8 Helper Free shRNA Expression System (Puro)

1 kit

VPK-412-DJ-8

AAV-DJ/8 Helper Free shRNA Expression System (GFP)

1 kit

VPK-413-DJ-8

scAAV-DJ/8 Helper Free Expression System

1 kit

VPK-430-DJ-8

Product Name

Size

Catalog Number

AAV-1 Helper Free Expression System

1 kit

VPK-410-SER1

AAV-1 Helper Free Bicistronic Expression System (Puro)

1 kit

VPK-415-SER1

AAV-1 Helper Free Bicistronic Expression System (Neo)

1 kit

VPK-416-SER1

AAV-1 Helper Free Bicistronic Expression System (Hygro)

1 kit

VPK-417-SER1

AAV-1 Helper Free Bicistronic Expression System (GFP)

1 kit

VPK-418-SER1

AAV-1 Helper Free Bicistronic Expression System (Blasticidin)

1 kit

VPK-419-SER1

AAV-1 Helper Free Promoterless Expression System

1 kit

VPK-411-SER1

AAV-1 Helper Free shRNA Expression System (Puro)

1 kit

VPK-412-SER1

AAV-1 Helper Free shRNA Expression System (GFP)

1 kit

VPK-413-SER1

scAAV-1 Helper Free Expression System

1 kit

VPK-430-SER1

Product Name

Size

Catalog Number

AAV-2 Helper Free Expression System

1 kit

VPK-410-SER2

AAV-2 Helper Free Bicistronic Expression System (Puro)

1 kit

VPK-415-SER2

AAV-2 Helper Free Bicistronic Expression System (Neo)

1 kit

VPK-416-SER2

AAV-2 Helper Free Bicistronic Expression System (Hygro)

1 kit

VPK-417-SER2

AAV-2 Helper Free Bicistronic Expression System (GFP)

1 kit

VPK-418-SER2

AAV-2 Helper Free Bicistronic Expression System (Blasticidin)

1 kit

VPK-419-SER2

AAV-2 Helper Free Promoterless Expression System

1 kit

VPK-411-SER2

AAV-2 Helper Free shRNA Expression System (Puro)

1 kit

VPK-412-SER2

AAV-2 Helper Free shRNA Expression System (GFP)

1 kit

VPK-413-SER2

scAAV-2 Helper Free Expression System

1 kit

VPK-430-SER2

AAV-1 Helper Free Complete Expression Systems

AAV-2 Helper Free Complete Expression Systems

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VIRAL EXPRESSION

AAV Expression

AAV-3 Helper Free Complete Expression Systems Product Name

Size

Catalog Number

AAV-3 Helper Free Expression System

1 kit

VPK-410-SER3

AAV-3 Helper Free Bicistronic Expression System (Puro)

1 kit

VPK-415-SER3

AAV-3 Helper Free Bicistronic Expression System (Neo)

1 kit

VPK-416-SER3

AAV-3 Helper Free Bicistronic Expression System (Hygro)

1 kit

VPK-417-SER3

AAV-3 Helper Free Bicistronic Expression System (GFP)

1 kit

VPK-418-SER3

AAV-3 Helper Free Bicistronic Expression System (Blasticidin)

1 kit

VPK-419-SER3

AAV-3 Helper Free Promoterless Expression System

1 kit

VPK-411-SER3

AAV-3 Helper Free shRNA Expression System (Puro)

1 kit

VPK-412-SER3

AAV-3 Helper Free shRNA Expression System (GFP)

1 kit

VPK-413-SER3

scAAV-3 Helper Free Expression System

1 kit

VPK-430-SER3

Product Name

Size

Catalog Number

AAV-4 Helper Free Expression System

1 kit

VPK-410-SER4

AAV-4 Helper Free Bicistronic Expression System (Puro)

1 kit

VPK-415-SER4

AAV-4 Helper Free Bicistronic Expression System (Neo)

1 kit

VPK-416-SER4

AAV-4 Helper Free Bicistronic Expression System (Hygro)

1 kit

VPK-417-SER4

AAV-4 Helper Free Bicistronic Expression System (GFP)

1 kit

VPK-418-SER4

AAV-4 Helper Free Bicistronic Expression System (Blasticidin)

1 kit

VPK-419-SER4

AAV-4 Helper Free Promoterless Expression System

1 kit

VPK-411-SER4

AAV-4 Helper Free shRNA Expression System (Puro)

1 kit

VPK-412-SER4

AAV-4 Helper Free shRNA Expression System (GFP)

1 kit

VPK-413-SER4

scAAV-4 Helper Free Expression System

1 kit

VPK-430-SER4

Product Name

Size

Catalog Number

AAV-5 Helper Free Expression System

1 kit

VPK-410-SER5

AAV-5 Helper Free Bicistronic Expression System (Puro)

1 kit

VPK-415-SER5

AAV-5 Helper Free Bicistronic Expression System (Neo)

1 kit

VPK-416-SER5

AAV-5 Helper Free Bicistronic Expression System (Hygro)

1 kit

VPK-417-SER5

AAV-5 Helper Free Bicistronic Expression System (GFP)

1 kit

VPK-418-SER5

AAV-5 Helper Free Bicistronic Expression System (Blasticidin)

1 kit

VPK-419-SER5

AAV-5 Helper Free Promoterless Expression System

1 kit

VPK-411-SER5

AAV-5 Helper Free shRNA Expression System (Puro)

1 kit

VPK-412-SER5

AAV-5 Helper Free shRNA Expression System (GFP)

1 kit

VPK-413-SER5

scAAV-5 Helper Free Expression System

1 kit

VPK-430-SER5

AAV-4 Helper Free Complete Expression Systems

AAV-5 Helper Free Complete Expression Systems

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AAV Expression

VIRAL EXPRESSION

AAV-6 Helper Free Complete Expression Systems Product Name

Size

Catalog Number

AAV-6 Helper Free Expression System

1 kit

VPK-410-SER6

AAV-6 Helper Free Bicistronic Expression System (Puro)

1 kit

VPK-415-SER6

AAV-6 Helper Free Bicistronic Expression System (Neo)

1 kit

VPK-416-SER6

AAV-6 Helper Free Bicistronic Expression System (Hygro)

1 kit

VPK-417-SER6

AAV-6 Helper Free Bicistronic Expression System (GFP)

1 kit

VPK-418-SER6

AAV-6 Helper Free Bicistronic Expression System (Blasticidin)

1 kit

VPK-419-SER6

AAV-6 Helper Free Promoterless Expression System

1 kit

VPK-411-SER6

AAV-6 Helper Free shRNA Expression System (Puro)

1 kit

VPK-412-SER6

AAV-6 Helper Free shRNA Expression System (GFP)

1 kit

VPK-413-SER6

scAAV-6 Helper Free Expression System

1 kit

VPK-430-SER6

AAV Helper Free Packaging Systems AAV Helper Free Packaging Systems contain everything found in the Complete Expression Systems, with the exception of the AAV expression vector. This is an ideal choice if you already have an AAV construct containing your gene of interest. All plasmids are provided individually, not as a packaging mixture. Product Name

Size

Catalog Number

AAV-DJ Helper Free Packaging System

1 kit

VPK-400-DJ

AAV-DJ/8 Helper Free Packaging System

1 kit

VPK-400-DJ-8

AAV-1 Helper Free Packaging System

1 kit

VPK-401

AAV-2 Helper Free Packaging System

1 kit

VPK-402

AAV-3 Helper Free Packaging System

1 kit

VPK-403

AAV-4 Helper Free Packaging System

1 kit

VPK-404

AAV-5 Helper Free Packaging System

1 kit

VPK-405

AAV-6 Helper Free Packaging System

1 kit

VPK-406

AAV Rep-Cap Plasmids (Serotype-Specific) AAV Rep-Cap plasmids allow you to make recombinant AAV of a specific serotype. These plasmids are ideal if you already have an AAV packaging system for a different serotype. Just substitute one of these plasmids into your AAV Helper Free Packaging System or Expression System. Product Name

Catalog Number

Product Name

Catalog Number

pAAV-DJ Vector

VPK-420-DJ

pAAV-DJ Vector

VPK-420-DJ

pAAV-DJ/8 Vector

VPK-420-DJ-8

pAAV-DJ/8 Vector

VPK-420-DJ-8

pAAV-RC1 Vector

VPK-421

pAAV-RC1 Vector

VPK-421

pAAV-RC2 Vector

VPK-422

pAAV-RC2 Vector

VPK-422

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VIRAL EXPRESSION

AAV Expression

AAV Expression Vectors Each of our AAV Expression Vectors may be used with any of our AAV Helper Free Systems, regardless of AAV serotype. Choose one of these vectors when you already have an AAV Packaging System but may want to use a different promoter or selection marker. Product Name

Recent Product Citation Sen, Y. et al. (2014). TDP-43 causes differential pathology in neuronal versus glial cells in the mouse brain. Human Mol. Genet. 10.1093/hmg/ddt662. (VPK-410)

Cloning Capacity

Size

Catalog Number

3 kb

10 µg

VPK-410

pAAV-IRES-Puro Expression Vector

1.8 kb

10 µg

VPK-415

pAAV-IRES-Neo Expression Vector

1.6 kb

10 µg

VPK-416

pAAV-IRES-Hygro Expression Vector

1.4 kb

10 µg

VPK-417

pAAV-IRES-GFP Expression Vector

1.7 kb

10 µg

VPK-418

pAAV-IRES-Bsd Expression Vector

2 kb

10 µg

VPK-419

pAAV-MCS Promoterless Expression Vector

3.9 kb

10 µg

VPK-411

pAAV-U6-Puro Expression Vector

2.2 kb

10 µg

VPK-412

pAAV-U6-GFP Expression Vector

2.1 kb

10 µg

VPK-413

pscAAV-MCS Expression Vector

1.5 kb

10 µg

VPK-430

pAAV-MCS Expression Vector

AAV Control Plasmids Choose one of our AAV control vectors when you already have an AAV Packaging System and want to make a transduction control virus. Product Name

Size

Catalog Number

pAAV-GFP Control Vector

10 µg

AAV-400

pAAV-Cre Control Vector

10 µg

AAV-401

pAAV-LacZ Control Vector

10 µg

AAV-402

pscAAV-GFP Control Vector

10 µg

AAV-410

Product Name

Size

Catalog Number

AAV1-GFP Control Virus

50 µL

AAV-301

AAV2 Null Control Virus

50 µL

AAV-300

AAV2-Cre Control Virus

50 µL

AAV-310

AAV2-GFP Control Virus

50 µL

AAV-302

AAV2-Luc Control Virus

50 µL

AAV-320

AAV3-GFP Control Virus

50 µL

AAV-303

AAV5-GFP Control Virus

50 µL

AAV-305

AAV6-GFP Control Virus

50 µL

AAV-306

AAV Premade Control Viruses All AAV premade viruses are provided at a concentration of 1 x 1012 GC/mL.

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AAV Expression

VIRAL EXPRESSION

293AAV Cell Line Our 293AAV cell line is selected from the parental 293 cell line for larger surface area, flattened morphology, and firmer attachment to culture plates, resulting in production of higher yields of AAV. Product Name

Size

Catalog Number

1 x 10 cells

293AAV Cell Line

AAV-100

ViraBind™ AAV Purification Kits Purification of AAV via ultracentrifugation can be tedious and time-consuming, and may result in low yields. ViraBind™ AAV Purification Kits use a one-step proprietary matrix followed by further purification and concentration using a centrifugal concentrator. The result is a higher AAV yield with high purity in a fraction of the time. Kits are suitable for AAV-2 or AAV-DJ; they will not work with other AAV serotypes.  High Purity: No contamination bands as seen on SDS gel  Fast Results: Obtain purified virus in about 3 hours  High Yields: Recovery rate >60%

Recent Product Citation Uchida, S. et al. (2010). Early life stress enhances behavioral vulnerability to stress through the activation of REST4-mediated gene transcription in the medial prefrontal cortex of rodents. J. Neurosci. 30:15007-15018. (VPK-140)

Purification Procedure for the ViraBind™ AAV Purification Kit.

Electrophoretic Profile of Purified AAV2-GFP.

Product Name

Capacity/Prep

Size

Catalog Number

ViraBind™ AAV Purification Kit

Two 10-cm dishes

10 Preps

VPK-140

2 Preps

VPK-141

ViraBind™ AAV Purification Mega Kit

Ten 15-cm dishes

10 Preps

VPK-141-5

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VIRAL EXPRESSION

AAV Expression

QuickTiter™ AAV Quantitation Kit Traditional AAV Quantitation by dot blot can be tedious, time consuming, and suffer from high inter-assay variability. Our QuickTiter™ AAV Quantitation Kit uses a proprietary technology to quantify AAV nucleic acid content of unpurified AAV-2 or AAV-DJ, or from purified AAV of any serotype.

 Fast Results: Obtain purified virus in less than 2 hours  High Sensitivity: Limit of detection 1 x 109 GC/mL from unpurified supernatant or 5 x 1010 GC/mL from purified AAV

200 175

150

RFU

125 100

50 25

0 0

100

125

150

AAV DNA STD (ng)

AAV-2 DNA Standard Curve. The QuickTiter™ AAV-2 DNA Standard was diluted as described in the assay protocol. Fluorescence was measured on a SpectraMax Gemini XS Fluorometer (Molecular Devices) with a 485/538 nm filter set and 530 nm cutoff.

Product Name QuickTiter™ AAV Quantitaiton Kit

Capacity/Prep

Size

Catalog Number

Fluorometric

20 Assays

VPK-145

ViraDuctin™ AAV Transduction Reagent Successful gene expression studies using AAV depend on high transduction efficiencies into host cells. Infection rates appear to be highest in S-phase cells, which can account for a very small fraction of a cell population. Our ViraDuctin™ AAV Transduction Reagent can significantly increase the transduction efficiency of AAV vectors in both dividing and non-dividing cells. Increases are greatest in non-dividing cells, but even cells in S-phase show a noticeable increase in transduction efficiencies.

 Higher Efficiencies: Significantly increase rate of infection of host cells  Low Toxicity: No noticeable effect on cell viability  Universal: Suitable for use with both dividing and non-dividing cells

Product Name ViraDuctin™ AAV Transduction Reagent

Size*

Catalog Number

10 Transductions

AAV-200

50 Transductions

AAV-201

*Number of transductions performed in 35mm culture dishes. May be modified for use in culture plates or larger dishes. See product insert.

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Adenoviral Expression

VIRAL EXPRESSION

Adenoviral Expression Kits & Reagents Recombinant adenoviruses are excellent tools for introducing genetic material into host cells, since they can infect a variety of mammalian cell types with high efficiency. They remain epichromosal upon infection, so they are only suitable for transient gene expression. We offer a complete workflow solution to your adenoviral expression studies:   

Viral Expression Systems Viral Packaging Cell Line Premade Recombinant Adenoviruses

  

Purification Kits Quantitation / Titer Kits Transduction Reagents

RAPAd® Adenoviral Expression Systems Compared to other adenoviral expression systems, RAPAd® Adenoviral Expression Systems produce recombinant adenovirus in a much shorter time (about 2-3 weeks) with a substantial reduction in wild-type adenovirus. The RAPAd systems use a backbone vector from which the 5’ ITR, packaging signal and E1 sequences have been removed. Additionally, serial amplification of the recombinant adenovirus does not increase the level of replicationcompetent adenovirus.  Virtually No Wild-Type Virus: Backbone vector engineered to produce <300 wild-type plaques per 109 particles, compared with 104-106 WT plaques per 109 VP with most other methods  Faster Production: Virus generated in 2-3 weeks compared to a few months with traditional methods  7 Complete Systems: Choose CMV or RSV for gene expression, EF-1 for miRNA expression, U6 for shRNA, or clone your own promoter along with your gene of interest using our Universal system

Adenovirus Production using the RAPAd® Adenoviral Expression System.

Recent Product Citations 1. Kothari, H. et al. (2010). Cystine 186-cystine 209 disulfide bond is not essential for the procoagulant activity of tissue factor or for its de-encryption. Blood 115:4273-4283. (VPK-252) 2. Li, P. et al. (2013). MicroRNA-638 is highly expressed in human vascular smooth muscle cells and inhibits PDGF-BBinduced cell proliferation and migration through targeting orphan nuclear receptor NOR1. Cardiovasc. Res. 10.1093/cvr/ cvt082. (VPK-253) Product Name

Promoter

Size

Catalog Number

RAPAd® Universal Adenoviral Expression System

None

1 Kit

VPK-250

RAPAd® RSV Adenoviral Expression System

RSV

1 Kit

VPK-251

RAPAd® CMV Adenoviral Expression System

CMV

1 Kit

VPK-252

RAPAd® Bicistronic Adenoviral Expression System (GFP)

CMV

1 Kit

VPK-254

RAPAd® miRNA Adenoviral Expression System

EF-1

1 Kit

VPK-253

RAPAd® shRNA Adenoviral Expression System (Puro)

1 Kit

VPK-255

RAPAd® shRNA Adenoviral Expression System (GFP)

1 Kit

VPK-256

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VIRAL EXPRESSION

Adenoviral Expression

293AD Cell Line for Adenoviral Packaging and Amplification The 293AD cell line is derived from the parental 293 cell line, but has been specifically selected for adenovirus applications and offers advantages over conventional 293 cells: flattened morphology, firm attachment to culture plates, and a larger surface area for superior transfection and greater viral yields.

Recent Product Citations 1. Peng, D. et al. (2011). Glutathione peroxidase 7 protects against oxidative DNA damage in oesophageal cells. Gut 61:1250-1260. (AD-100) 2. Kothari, H. et al. (2010). Cystine 186-cystine 209 disulfide bond is not essential for the procoagulant activity of tissue factor or for its de-encryption. Blood 115:4273-4283. (AD-100)

Product Name

Size

Catalog Number

AD-100

1 x 10 Cells

293AD Cell Line

Premade Recombinant Adenoviruses Angiogenesis

Don’t have time to make your own adenovirus? Are you studying the expression of multiple genes? Rely on our premade recombinant adenoviruses that already contain a gene of interest. All of Cell Biolabs’ premade recombinant adenoviruses are provided as 50 µl aliquots at a concentration of 1 x 1011 viral particles/mL in TBS with 10% glycerol.

Controls and Reporter Genes Recent Product Citations 1. Matsushita, T. et al. (2009). FGFR3 promotes synchondosis closure and fusion of ossification centers through the MAPK pathway. Hum. Mol. Genet. 18:227-240. (ADV-001) 2. Zhang, Z. et al (2013). MEK inhibition leads to lysosomemediated Na+/I– symporter protein degradation in human breast cancer cells. Endocr. Relat. Cancer 20:241-250. (ADV002) 3. Schramm, C. et al (2012). The PTPN11 loss-of-function mutation Q510E-Shp2 causes hypertrophic cardiomyopathy by dysregulating mTOR signaling. Am. J. Physiol. Heart Circ. Physiol. 302:H231-H243. (ADV-002) 4. Salvati, E. et al. (2014). Evidence for G-quadruplex in the promoter of vegfr-2 and its targeting to inhibit tumor angiogenesis. Nucleic Acids Res. 42:2945-2957. (ADV-004) 5. Lu, D. et al. (2012). Peroxisome proliferator-activated receptorcoactivator-1alpha enhances engraftment and angiogenesis of mesenchymal stem cells in diabetic hindlimb ischemia. Diabetes 61:1153-1159. (ADV-004) 6. Kato, H. et al. (2011). Wnt/ß-Catenin pathway in podocytes integrates cell adhesion, differentiation and survival. J. Biol. Chem. 286:26003-26015. (ADV-005)

Target Name

Catalog Number

Blood Vessel Formation After 3 Days. Purified Ad-Null or AdVEGF viruses were applied to a 10-day old CAM (chick chorioallanoic membrane). Results were visualized by stereomicroscope. Recent Product Citations 1. Kelber, J.A. et al. (2012). KRas induces a Src/PEAK1/ErbB2 kinase amplification loop that drives metastatic growth and therapy resistance in pancreatic cancer. Cancer Res. 72:25542564. (ADV-101) 2. Qiu, X. et al. (2012). Combined strategy of mesenchymal stem cell injection with vascular endothelial growth factor gene therapy for the treatment of diabetes-associated erectile dysfunction. J. Androl. 33:37-44. (ADV-101) 3. Stoletov, K. et al. (2010). Visualizing extravasation dynamics of metastatic tumor cells. J. Cell Sci. 123:2332-2341. (ADV-101) 4. Serban, D. et al. (2008). H-ras regulates angiogenesis and vascular permeability by activation of distinct downstream effectors. Circ. Res. 102(11):1350-1358. (ADV-101) Target Name

Catalog Number

HIF-1

ADV-100

VEGF

ADV-101

Null Control (No gene)

ADV-001

-Galactosidase

ADV-002

Cre

ADV-005

Target Name

Firefly Luciferase

ADV-008

Carbonic Anhydrase 9 (CA9)

ADV-602

GFP

ADV-004

Carcinoembryonic Antigen (CEA)

ADV-604

SEAP (Secretory Alkaline Phosphatase)

ADV-003

NY-ESO-1

ADV-601

Phone 1-858-271-6500

Cancer/Tumor Antigens

USA Toll-Free 1-888-CBL-0505

Catalog Number

Fax 1-858-271-6514

Adenoviral Expression

VIRAL EXPRESSION

Premade Recombinant Adenoviruses, continued Cell Cycle & Transcription Regulation Recent Product Citation Nguyen, N. et al. (2014). Mitsugumin 53 (MG53) ligase ubiquitinates focal adhesion kinase during skeletal myogenesis. J. Biol. Chem. 289:3209-3216. (ADV-508)

Myogenin

ADV-509

Target Name

p53

ADV-501

Catalog Number

Target Name

Catalog Number

DCC

ADV-504

p53 (Temperature Sensitive Mutant)

ADV-502

MyoD

ADV-508

p68 RNA Helicase

ADV-505

Cytoskeleton Regulation / Small GTPase

Cdc42

ADV-152

Cdc42 L61 (Constitutively Active)

ADV-154

Cdc42 N17 (Dominant Negative)

ADV-153

PAK1

ADV-202

PAK1 (H83L, H86L)

ADV-203

PAK1 (H83L, H86L, K299R)

ADV-205

Recent Product Citations 1. Black, S.A. et al. (2008). TGFß1 stimulates connective tissue growth factor (CCN2/CTGF) expression in human gingival fibroblasts through a RhoA-independent, Rac1/Cdc42dependent mechanism: statins with forskolin block TGFß1induced CCN2/CTGF expression. J. Biol. Chem. 283:1083510847. (ADV-145, ADV-150, ADV-153, ADV-156) 2. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires vß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Cell Biol. 23:1104-1114. (ADV-150) 3. Thomas, M.A. et al. (2009). E4orf1 limits the oncolytic potential of the E1B-55K-deleted adenovirus. J. Virol. 83:2406-2416. (ADV-150) 4. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150, ADV-153, ADV-154) 5. Salvati, E. et al. (2014). Evidence for G-quadruplex in the promoter of vegfr-2 and its targeting to inhibit tumor angiogenesis. Nucleic Acids Res. 42:2945-2957. (ADV-151, ADV-157) 6. Cheng, Z.-J. et al. (2010). Co-regulation of caveolar and Cdc42 -dependent fluid phase endocytosis by phosphocaveolin-1. J. Biol. Chem. 285:15119-15125. (ADV-153) 7. Neal M. et al. (2013). A critical role for TLR4 induction of autophagy in the regulation of enterocyte migration and the pathogenesis of necrotizing enterocolitis. J. Immunol. 190:35413551. (ADV-156, ADV-157) 8. Nie, J. et al. (2013). SAD-A kinase controls islet ß-cell size and function as a mediator of mTORC1 signaling. PNAS 110:13857 -13862. (ADV-204, ADV-207) 9. Nie, J. et al. (2013). Synapses of amphids defective (SAD-A) kinase promotes glucose-stimulated insulin secretion through activation of p21-activated kinase (PAK1) in pancreatic ß-cells. J. Biol. Chem. 287:26435-26444. (ADV-204, ADV-207)

PAK1 (K299R)

ADV-207

Target Name

PAK1 (L107E, T423E)

ADV-206

Ras N17 (Dominant Negative)

ADV-145

PAK1 (T423E)

ADV-204

Ras V12 (Constitutively Active)

ADV-146

PAK1 (Kinase Domain)

ADV-209

Ras V12C40

ADV-148

PAK1 (Regulatory Domain)

ADV-208

Ras V12S35

ADV-147

Rac1

ADV-149

Rho L63 (Constitutively Active)

ADV-157

Rac1 L61 (Constitutively Active)

ADV-151

Rho N19 (Dominant Negative)

ADV-156

Rac1 N17 (Dominant Negative)

ADV-150

SDF-1

ADV-210

Actin Cytoskeleton Staining. Cos-7 cells were infected with purified Ad-Ras V12 (ADV-146) at 50 MOI (multiplicity of infection). Membrane ruffling was visualized by staining the actin cytoskeleton with Rhodamine-coupled Phalloidin. Target Name

Catalog Number

www.cellbiolabs.com

Catalog Number

info@cellbiolabs.com

VIRAL EXPRESSION

Adenoviral Expression

Premade Recombinant Adenoviruses, continued MAP Kinase Signaling Immunoblot of Various Cell Signaling Targets. HUVEC cells were infected with purified AdNull (ADV-001), Ad-Ras V12 (ADV-146), Ad-Ras V12S35 (ADV-147), AdRas V12C40 (ADV-148), Ad-MEK1 (ADV-119), and Ad-Rac1 L61 (ADV151) at 10 MOI (multiplicity of infection). Cell lysates were analyzed for gene expression and ERK activation.

See following page for recent citations using these adenoviruses Target Name

Catalog Number

Target Name

Catalog Number

MKK3

ADV-120

MKK3 (Dominant Negative)

ADV-121

MKK3 (Constitutively Active)

ADV-122

MKK4 (Dominant Negative)

ADV-160

MKK4 (Constitutively Active)

ADV-161

MKK6

ADV-123

MKK6 (Dominant Negative)

ADV-124

MKK6 (Constitutively Active)

ADV-125

MKK7

ADV-126

MKK7 (Dominant Negative)

ADV-127

MKK7 (Constitutively Active)

ADV-128

myr-Rac1

ADV-163

Cdc42

ADV-152

p38

ADV-104

Cdc42 L61 (Constitutively Active)

ADV-154

p38 (Dominant Negative)

ADV-105

Cdc42 N17 (Dominant Negative)

ADV-153

p38

ADV-106

ERK2

ADV-112

p38 (Dominant Negative)

ADV-107

ERK2 (Dominant Negative)

ADV-113

p38

ADV-108

ERK5 (BMK1)

ADV-116

p38 (Dominant Negative)

ADV-109

ERK5 (Dominant Negative)

ADV-117

p38 (Dominant Negative)

ADV-111

Interferon-

ADV-103

PRAK (Dominant Negative)

ADV-141

Interleukin-2

ADV-102

Rac1

ADV-149

JNK1

ADV-114

Rac1 L61 (Constitutively Active)

ADV-151

JNK1 (Dominant Negative)

ADV-115

Rac1 N17 (Dominant Negative)

ADV-150

MAPKAPK2

ADV-137

Raf1

ADV-132

MAPKAPK2 (Dominant Negative)

ADV-138

Raf1 (Dominant Negative)

ADV-133

MAPKAPK2 (Constitutively Active)

ADV-139

Raf1 (Constitutively Active)

ADV-134

MEK1 (Dominant Negative)

ADV-118

Ras N17 (Dominant Negative)

ADV-145

MEK1 (Constitutively Active)

ADV-119

Ras V12 (Constitutively Active)

ADV-146

MEK5

ADV-129

Rho L63 (Constitutively Active)

ADV-157

MEK5 (Dominant Negative)

ADV-130

Rho N19 (Dominant Negative)

ADV-156

MEK5 (Constitutively Active)

ADV-131

SOK

ADV-142

MEKK1

ADV-135

SOK (Dominant Negative)

ADV-143

MEKK1 (Dominant Negative)

ADV-136

SOK (Constitutively Active)

ADV-144

MEKK3

ADV-162

Tac-Rac1 (Membrane Targeting)

ADV-164

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Adenoviral Expression

VIRAL EXPRESSION

Premade Recombinant Adenoviruses, continued MAP Kinase Signaling, continued Recent Product Citations 1. Harbrecht, B.G. et al. (2012). Insulin inhibits hepatocyte iNOS expression induced by cytokines by an Akt-dependent mechanism. Am. J. Physiol. Gastrointest. Liver Physiol 302:G116G122. (ADV-105) 2. Jones, S.W. et al. (2009). Mitogen-activated protein kinaseactivated protein kinase (MK2) modulates key biological pathways associated with OA disease pathology. Osteoarthritis and Cartilage 17:124-131. (ADV-105) 3. Kim, J.M. et al. (2008). Inhibition of apoptosis in Bacteroids fragilis enterotoxin-stimulated intestinal epithelial cells through the induction of c-IAP-2. Eur. J. Immunol. 38(8):2190-2199. (ADV-105) 4. Lee, J.Y. et al. (2007). Effects of transcription factor activator protein-1 on interleukin-8 expression and enteritis in response to Clostridium difficile toxin A. J. Mol. Med. 85:1393-1404. (ADV-105, ADV-115) 5. Monick, M. et al. (2008). Constitutive ERK MAPK activity regulates macrophage ATP production and mitochondrial integrity. J. Immunol. 180:7485-7496. (ADV-112, ADV-113, ADV-118, ADV-119) 6. Samuel, I. et al. (2008). Enteral exclusion increases MAP kinase activation and cytokine production in a model of gallstone pancreatitis. Pancreatology 8(1):6-14. (ADV-113) 7. Wang, X. et al. (2007). Human immunodeficiency virus protease inhibitor ritonavir inhibits cholesterol efflux from human macrophage-derived foam cells. Am. J. of Pathology 171:304314. (ADV-113) 8. Jiang, S. et al. (2011). Role of inhibitory kB kinase and c-Jun NH2-terminal kinase in the development of hepatic insulin resistance in critical illness diabetes. Am. J. Physiol. Gastrointest. Liver Physiol 301:G454-G463. (ADV-115) 9. Zhang, Z. et al. (2013). MEK inhibition leads to lysosomemediated Na+/I– symporter protein degradation in human breast cancer cells. Endocr. Relat. Cancer 20:241-250. (ADV118) 10. Matsushita, T. et al. (2009). FGFR3 promotes synchondosis closure and fusion of ossification centers through the MAPK pathway. Hum. Mol. Genet. 18:227-240. (ADV-119) 11. Yoon, C-H. et al. (2009). Activation of p38 mitogen-activated protein kinase is required for death receptor-independent caspase-8 activation and cell death in response to sphingosine. Mol. Cancer Res. 7(3):361-370. (ADV-119) 12. Tan, S.H. et al. (2009). Regulation of cell proliferation and migration by TAK1 via transcriptional control of von Hippel-Lindau tumor suppressor. J. Biol. Chem. 284:18047-18058. (ADV-128) 13. Wu, Y. et al. (2012). ERK5 regulates glucose-induced increased fibronectin production in the endothelial cells and in the retina in diabetes. Invest. Ophthalmol. Vis. Sci. 53:84058413. (ADV-130) 14. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires vß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Cell Biol. 23:1104-1114. (ADV-150) 15. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150) 16. Neal, M. et al. (2013). A critical role for TLR4 induction of autophagy in the regulation of enterocyte migration and the pathogenesis of necrotizing enterocolitis. J. Immunol. 190:35413551. (ADV-156, ADV-157) 17. Taniguchi, C. et al. (2007). The p85a regulatory subunit of phosphoinositide 3-kinase potentiates c-Jun N-terminal kinasemediated insulin resistance. Mol. Cell Biol. 27:2830-2840. (ADV-161)

NFB Signaling Recent Product Citations 1. Johnston, R.K. et al. (2009). ß3-integrin mediated ubiquitination activates survival signaling during myocardial hypertrophy. FASEB J. 23(8):2759-2771. (ADV-302) 2. Martin, A.P. et al. (2008). Lapatinib resistance in HCT116 cells is mediated by elevated MCL-1 expression and decreased BAK activation and not by ERBB receptor kinase mutation. Mol. Pharmacol. 74:807-822. (ADV-302) 3. Richardson, W.M. et al. (2010). Nucleotide-binding oligomerization domain-2 inhibits toll-like receptor-4 signaling in the intestinal epithelium. Gastroenterology 139(3):904-917. (ADV308, ADV-309) Target Name

Catalog Number

IB-

ADV-301

IB- S32A (Dominant Negative)

ADV-302

IKK-

ADV-305

IKK- (Dominant Negative)

ADV-303

NOD2

ADV-308

Rel B

ADV-304

Tyrosine Kinases and PKCs Recent Product Citations 1. Koh, W. et al. (2009). Formation of endothelial lumens requires a PKC-, Src-, Pak-, and Raf-kinase dependent signaling cascade downstream of Cdc42 activation. J. Cell Sci. 122:1812-1822. (ADV -401, ADV-405, ADV-406) 2. Lecuona, E. et al. (2009). Ubiquitination participates in the lysosomal degradation of the Na,K-ATPase in steady state conditions. Am. J. Respir. Cell Mol. Biol. 41(6):617-679. (ADV-412) 3. Vadasz, I. et al. (2008). AMP-activated protein kinase regulates CO2-induced alveolar epithelial dysfunction in rats and human cells by promoting Na,K-ATPase endocytosis. J. Clin. Invest. 118 (2):752-762. (ADV-412) 4. Briva, A. et al. (2007). High CO2 levels impair alveolar epithelial function independent of pH. PLoS ONE 2(11):e1238. (ADV-412) Target Name

Catalog Number

CSK

ADV-405

CSK (Dominant Negative)

ADV-406

Fyn

ADV-403

Fyn (Dominant Negative)

ADV-404

PKC- (Dominant Negative)

ADV-410

PKC- (Dominant Negative)

ADV-411

PKC- (Dominant Negative)

ADV-412

shAkt1

ADV-417

shAkt2

ADV-418

Src

ADV-401

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VIRAL EXPRESSION

Adenoviral Expression

ViraBind™ Adenovirus Purification Kits Purification of viruses via cesium chloride (CsCl) ultracentrifugation procedures can be tedious and timeconsuming. ViraBind™ Adenovirus Purification Kits use an efficient system for quick adenoviral purification with high recovery. No ultracentrifugation is required. Kits use either a spin column or syringe filter for high purity adenovirus (see selection guide).  High Viral Yield: >90% recovery  High Quality: Provides quality of CsCl procedures, but in much less time  Faster Results: 30 minutes (1-2 hrs for Mega kit)  User-Friendly Protocol: No gradient preparation or ultracentrifugation steps

Relative VP (%)

100 80 60 40 20

El ut io n

W as

tW as h 1s

gh ro u

Fl ow - th

Vi ra l

pe rn a

ta n

Purification of Recombinant Ad--Gal. Ad-β-Gal was purified according to the assay protocol. Each purification fraction was used to infect A549 cells in a 12-well plate. After 48 hr, cells were scored using our β-Galactosidase Staining Kit (p. 92).

Recent Product Citations 1. Wilkins, H. et al. (2013). Mitochondrial glutathione transport is a key determinant of neuronal susceptibility to oxidative and nitrosative stress. J. Biol. Chem. 288:5091-5101. (VPK-099) 2. Wang, Y.S. et al. (2012). MicroRNA-195 regulates vascular smooth muscle cell phenotype and prevents neointimal formation. Cardiovasc. Res. 95:517-526. (VPK-099) 3. Kirui, J.K. et al. (2010). Gßgamma signaling promotes breast cancer cell migration and invasion. J. Pharmacol. Exp. Ther. 333:393-403. (VPK-099) 4. Scallan, C. et al. (2013). An adenovirus-based vaccine with a double-stranded RNA adjuvant protects mice and ferrets against H5N1 avian influenza in oral delivery models. Clin. Vaccine Immunol. 20:85-94. (VPK-100) 5. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phosphorylation of vascular endothelial cadherin induced by invasive breast cancer cells. J. Biol. Chem. 287:32981-32992. (VPK-100) 6. Chen, F. et al. (2011). Dynamic regulation of PDX-1 and FoxO1 expression by FoxA2 in dexamethasone-induced pancreatic ß-cells dysfunction. Endocrinology 152:1779-1788. (VPK-100) 7. Prasad, S.S. et al. (2011). Enzymatic activities of the human AGPAT isoform 3 and isoform 5: localization of AGPAT5 to mitochondria. J. Lipid Res. 52:451-462. (VPK-100) 8. Agarwal, A.K. et al. (2010). Enyzmatic activity of the human 1acylglycerol-3-phosphate-O-acyltransferase isoform 11: upregulated in breast and cervical cancers. J. Lipid Res. 51:2143-2152. (VPK-100) 9. Triulzi, C. et al. (2010). Antibody-dependent natural killer cellmediated cytotoxicity engendered by a kinase-inactive HER2 adenovirus-based vaccination mediates resistance to breast tumors. Cancer Res. 70:7431-7441. (VPK-100) 10.Sen, P. et al. (2010). Zinc modulates the interaction of protein C and activated protein C with endothelial protein C receptor. J. Biol. Chem. 285:20410-20420. (VPK-100) 11.Sabbatini, M. E. et al. (2010). CCK activates RhoA and Rac1 differentially through G-alpha-13 and G-alpha-q in mouse pancreatic acini. Am. J. Physiol. Cell Physiol. 298:C592-C605. (VPK-100) 12.Lam, Y.W. et al. (2010). Proteomics analysis of the nucleolus in adenovirus-infected cells. Mol. Cell. Proteomics 9:117-130. (VPK-100)

Selection Guide for ViraBind™ Adenovirus Purification Kits

Purification Method Purification Time Capacity/Prep (Viral Particles) Capacity/Prep (Supernatant)

Product Name ViraBind™ Adenovirus Miniprep Kit ViraBind™ Adenovirus Purification Kit

Phone 1-858-271-6500

ViraBind™ Adenovirus Miniprep Kit

ViraBind™ Adenovirus Purification Kit

Spin column

Syringe filter

30 minutes

1 x 10 VP

2.5 x 1012 VP

One T75 flask or one 10cm dish

Four T75 flasks

Capacity/Prep 1 x 10

2.5 x 10

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

10 Preps

VPK-099

10 Preps

VPK-100

Fax 1-858-271-6514

Adenoviral Expression

VIRAL EXPRESSION

QuickTiter™ Adenoviral Titer & Quantitation Kits Accurate measurement of virus titer is critical for viral gene delivery. Traditional plaque-forming unit (PFU) assays are long and have high inter-assay variability. The QuickTiter™ Adenovirus Titer Kits provide a complete system to functionally titer virus infectivity with greater accuracy in a fraction of the time. The assays may be used with any adenovirus system that can amplify in 293 cells. Assays are available for ICC staining or 96-well ELISA. For a quick test of physical titer, our QuickTiter™ Adenovirus Quantitation Kit measures the concentration of your adenovirus prep in about one hour.

QuickTiter™ Adenovirus Titer Immunoassay Kit. 293AD cells (p. 42) were infected with different dilutions of purified Ad-β-Gal for 48 hours. Immunostaining was performed according to the assay protocol. X-gal staining was performed with β-Galactosidase Staining Kit (p. 106).

 Faster, More Accurate and Precise: Compared to traditional plaque-forming unit assays  User-Friendly Protocol: No agar overlay steps  Versatile: Recognize all 41 adenovirus serotypes Recent Product Citations 1. Smith, M. et al. (2010). PRDM1/Blimp-1 controls effector cytokine production in human NK cells. J. Immunol. 185:6058-6067. (VPK-106) 2. Reiter, C.E.N. et al. (2010). Green tea polyphenol epigallocatechin gallate reduces endothelin-1 expression and secretion in vascular endothelial cells: roles for AMP-activated protein kinase, Akt, and FOXO1. Endocrinology 151:103-114. (VPK106) 3. Wilkins, H. et al. (2013). Mitochondrial glutathione transport is a key determinant of neuronal susceptibility to oxidative and nitrosative stress. J. Biol. Chem. 288:5091-5101. (VPK-109) 4. Scallan, C. et al. (2013). An adenovirus-based vaccine with a double-stranded RNA adjuvant protects mice and ferrets against H5N1 avian influenza in oral delivery models. Clin. Vaccine Immunol. 20:85-94. (VPK-109) 5. Xiong, X. et al. (2012). The autophagy-related gene 14 (Atg14) is regulated by forkhead box O transcription factors and circadian rhythms and plays a critical role in hepatic autophagy and lipid metabolism. J. Biol. Chem. 287:39107-39114. (VPK109) 6. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phosphorylation of vascular endothelial cadherin induced by invasive breast cancer cells. J. Biol. Chem. 287:32981-32992. (VPK-109) 7. Hisamitsu, T. et al. (2012). Na+/H+ exchanger 1 directly binds to calcineurin A and activates downstream NFAT signaling, leading to cardiomyocyte hypertrophy. Mol. Cell Biol. 32:32653280. (VPK-109) 8. Lee, S. et al. (2012). Adiponectin abates diabetes-induced endothelial dysfunction by suppressing oxidative stress, adhesion molecules, and inflammation in type 2 diabetic mice. Am. J. Heart Circ. Physiol. 303:H106-H115. (VPK-109)

Selection Guide for QuickTiter™ Adenoviral Quantitation Kits

Functional or Physical Titer

QuickTiter™ Adenovirus Titer Immunoassay Kit

QuickTiter™ Adenovirus Titer ELISA Kit

QuickTiter™ Adenovirus Quantitation Kit

Functional (Infectious units)

Physical (Viral particles)

2.5 days

45-60 minutes

Antibody-based

Total nucleic acid content

Immunocytochemical staining

Colorimetric (ELISA) plate reader

Fluorescence plate reader

Accuracy

Speed

Assay Time Assay Principle Detection Method Key Benefit Product Name

Detection

Size

Catalog Number

QuickTiter™ Adenovirus Titer Immunoassay Kit

ICC Staining

100 Assays

VPK-109

QuickTiter™ Adenovirus Titer ELISA Kit

Colorimetric

2 x 96 Assays

VPK-110

QuickTiter™ Adenovirus Quantitation Kit

Fluorometric

20 Assays

VPK-106

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VIRAL EXPRESSION

Adenoviral Expression

Rapid Replication Competent Adenovirus (RCA) Assay Kit This kit uses the assay principles of the QuickTiter™ Adenovirus Titer Immunoassay Kit (see page 47), but is designed specifically to measure the level of replication-competent virus in your adenoviral prep.

Immunostaining of Wild Type Ad5 using the Rapid RCA Assay Kit.

 Faster Results: 2.5 days vs. 10 days with plaque assay  Versatile: Recognizes all 41 adenovirus serotypes Product Name

Detection

Rapid RCA Assay Kit

ICC Staining

Size

Catalog Number

30 Assays

VPK-111

5 x 30 Assays

VPK-111-5

ViraDuctin™ Adenovirus Transduction Reagent, CAR-Independent Adenovirus infection of target cells is mediated largely by the coxsackievirus-adenovirus receptor (CAR). Generally adenoviral transduction of many immortalized cell lines proceeds with a high level of efficiency. However, in many primary cells this receptor is either absent or present at extremely lowlevels. This can reduce the efficiency of adenovirus transduction into your cell of choice.

 Higher Transduction Efficiency: Up to 12-fold increase in adenoviral uptake  User-Friendly: Short incubation step prior to host cell infection  Versatile: Ideal for target cells expressing little or no CAR, but may also improve transduction efficiency for CAR-expressing cells Recent Product Citations 1. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phosphorylation of vascular endothelial cadherin induced by invasive breast cancer cells. J. Biol. Chem. 287:32981-32992. 2. Ackerman, W. et al (2008). Nuclear Factor-kappa B regulates inducible prostaglandin E synthase expression in human amnion mesenchymal cells. Biol. Reprod. 78:68-76. 3. Monick, M. et al. (2008). Constitutive ERK MAPK activity regulates macrophage ATP production and mitochondrial integrity. J. Immunol. 180:7485-7496.

1400 X-Gal Staining Positive (%)

ViraDuctin™ Adenovirus Transduction Reagent is designed specifically to increase the efficiency of adenoviral transduction, without regard to the level of CAR expression on the surface of the target cells.

1200 1000 800 600 400 200 0 Control

ViraDuctin™

Enhanced Transduction using ViraDuctin™ Adenovirus Transduction Reagent. Infection of NIH3T3 cells with recombinant Ad-ß -gal (ADV-002). Top: X-gal staining under microscope. Bottom: scoring of infection with ViraDuctin™ reagent as a percentage of infection with control.

Product Name ViraDuctin™ Adenovirus Transduction Reagent (CAR-Independent)

Size*

Catalog Number

10 Transductions

AD-200

50 Transductions

AD-201

*Based on using 6-well plates or 35mm culture dishes; may also be used with 96-,24- or 12-well plates or 60mm or 100mm dishes.

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Lentiviral Expression

VIRAL EXPRESSION

Lentiviral Expression Kits & Reagents As a sub-class of retroviruses, lentiviruses based on HIV-1 have the unique advantage of being able to infect both proliferating and non-proliferating cells, and they can be used for both transient and stable gene expression. We offer a complete workflow solution to your lentiviral expression studies:   

Expression Systems & Vectors Premade Controls Viral Host Cell Line

  

Concentration / Purification Kits Quantitation / Titer Kits Transduction Reagents

ViraSafe™ Lentiviral Expression Systems Lentiviruses based on HIV-1 may infect both dividing and non-dividing cells. Recently developed thirdgeneration lentiviral expression systems have reduced the risk of creating replication-competent virus upon recombination, but the risk is still present. Our ViraSafe™ Lentiviral Expression Systems provide a safer and more flexible method to package your lentivirus, even compared to other thirdgeneration lentivirus systems.  Safer: 80-90% less sequence homology compared to other 3rd-generation lentiviral systems; ecotropic systems provide even more safety*  High Titers: Incorporates elements that provide titers comparable to other 3rd-generation systems  Flexible: Packaging vectors provided separately for increased safety and optimization of vector ratios

*Lentiviruses made with a ViraSafe™ Ecotropic Expression System will only readily infect mouse and rat cells. Pantropic lentiviruses are VSVG-pseudotyped and may infect cells of any species. ViraSafe™ Lentiviral Technology is available in three formats (see next two pages for ordering information):  Complete Expression Systems: Include 3 packaging plasmids, expression vector and control vector  Packaging Systems: Include the 3 individual packaging plasmids; ideal if you already have a 3rdgeneration lentiviral expression construct  Expression Vectors: 11 cloning vectors to choose from; compatible with any 2nd or 3rd generation packaging system, but produce the highest titers with the ViraSafe™ packaging system

Lentivirus Production using the ViraSafe™ Lentiviral Expression System.

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VIRAL EXPRESSION

Lentiviral Expression

ViraSafe™ Lentiviral Expression Systems, continued Complete ViraSafe™ Expression Systems include three individual packaging plasmids, an expression vector, and a control vector. Choose an ecotropic system for infection of mouse or rat cells, or a pantropic system to produce VSVG-pseudotyped lentivirus for infection of cells from any species.

Recent Product Citation Davis, M. et al. (2013). RAC1P29S is a spontaneously activating cancer-associated GTPase. PNAS 110:912-917. (VPK-214-PAN)

Product Name ViraSafe™ Universal Lentiviral Expression System (Promoterless)

ViraSafe™ Lentiviral Expression System (Puro)

ViraSafe™ Lentiviral Expression System (Neo)

ViraSafe™ Lentiviral Expression System (Hygro)

ViraSafe™ Lentiviral Bicistronic Expression System (Puro)

ViraSafe™ Lentiviral Bicistronic Expression System (Neo)

ViraSafe™ Lentiviral Bicistronic Expression System (Hygro)

ViraSafe™ Lentiviral Bicistronic Expression System (GFP)

ViraSafe™ Lentiviral Bicistronic Expression System (Blasticidin)

ViraSafe™ shRNA Lentiviral Expression System (Puro)

ViraSafe™ shRNA Lentiviral Expression System (GFP)

Envelope

Size

Catalog Number

Ecotropic

1 Kit

VPK-211-ECO

Pantropic (VSVG)

1 Kit

VPK-211-PAN

Ecotropic

1 Kit

VPK-212-ECO

Pantropic (VSVG)

1 Kit

VPK-212-PAN

Ecotropic

1 Kit

VPK-213-ECO

Pantropic (VSVG)

1 Kit

VPK-213-PAN

Ecotropic

1 Kit

VPK-214-ECO

Pantropic (VSVG)

1 Kit

VPK-214-PAN

Ecotropic

1 Kit

VPK-215-ECO

Pantropic (VSVG)

1 Kit

VPK-215-PAN

Ecotropic

1 Kit

VPK-216-ECO

Pantropic (VSVG)

1 Kit

VPK-216-PAN

Ecotropic

1 Kit

VPK-217-ECO

Pantropic (VSVG)

1 Kit

VPK-217-PAN

Ecotropic

1 Kit

VPK-218-ECO

Pantropic (VSVG)

1 Kit

VPK-218-PAN

Ecotropic

1 Kit

VPK-219-ECO

Pantropic (VSVG)

1 Kit

VPK-219-PAN

Ecotropic

1 Kit

VPK-221-ECO

Pantropic (VSVG)

1 Kit

VPK-221-PAN

Ecotropic

1 Kit

VPK-222-ECO

Pantropic (VSVG)

1 Kit

VPK-222-PAN

ViraSafe™ Lentiviral Packaging Systems ViraSafe™ Packaging Systems contain 3 packaging plasmids for use with any 3rd-generation lentiviral expression vector. These systems are perfect if you already have a lentiviral construct containing your gene of interest. Product Name ViraSafe™ Lentiviral Packaging System

Phone 1-858-271-6500

Recent Product Citation Vogt, J. et al. (2014). Protein associated with SMAD1 (PAWS1/ FAM83G) is a substrate for type I bone morphogenetic protein receptors and modulates bone morphogenetic protein signaling. Open Bio. 4:130210. (VPK-206)

Envelope

Size

Catalog Number

Ecotropic

1 Kit

VPK-205

Pantropic (VSVG)

1 Kit

VPK-206

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

VIRAL EXPRESSION

Lentiviral Expression ViraSafe™ Lentiviral Expression Vectors These lentiviral expression vectors may be used with any 2nd or 3rd generation lentiviral packaging system, but best results are achieved when used with our ViraSafe™ Lentiviral Packaging Systems. Product Name

Recent Product Citations 1. Sankaran, V.G. et al. (2011). MicroRNA-15a and -16-1 act via MYB to elevate hemoglobin expression in human trisomy 13. PNAS 108(4):1519-1524. (VPK-212) 2. Davis, M. et al. (2013). RAC1P29S is a spontaneously activating cancer-associated GTPase. PNAS 110:912-917. (VPK-214) Cloning Capacity

Size

Catalog Number

pSMPUW Universal Lentiviral Expression Vector (Promoterless)

9.4 kb

10 µg

VPK-211

pSMPUW-Puro Lentiviral Expression Vector

7.9 kb

10 µg

VPK-212

pSMPUW-Neo Lentiviral Expression Vector

7.7 kb

10 µg

VPK-213

pSMPUW-Hygro Lentiviral Expression Vector

7.4 kb

10 µg

VPK-214

pSMPUW-IRES-Puro Lentiviral Expression Vector

7.8 kb

10 µg

VPK-215

pSMPUW-IRES-Neo Lentiviral Expression Vector

7.5 kb

10 µg

VPK-216

pSMPUW-IRES-Hygro Lentiviral Expression Vector

7.3 kb

10 µg

VPK-217

pSMPUW-IRES-GFP Lentiviral Expression Vector

7.6 kb

10 µg

VPK-218

pSMPUW-IRES-Bsd Lentiviral Expression Vector

7.9 kb

10 µg

VPK-219

pSMPUW-U6-Puro Lentiviral Expression Vector

7.7 kb

10 µg

VPK-221

pSMPUW-U6-GFP Lentiviral Expression Vector

7.6 kb

10 µg

VPK-222

Product Name

Size

Catalog Number

pLenti-GFP Lentiviral Control Vector

10 µg

LTV-400

pSMPUW-GFP-Puro Lentiviral Control Vector

10 µg

LTV-401

pSMPUW-MNDnLacZ Lentiviral Control Vector

10 µg

LTV-402

pLenti-RFP-Puro Lentiviral Control Vector

100 µL

LTV-403

Lentiviral Control Plasmids

Premade Reporter Lentivirus Controls Product Name GFP Lentivirus Control RFP Lentivirus Control

Concentration

Size

Catalog Number

200 µL

LTV-300

200 µL

LTV-301

1 x 10 TU/mL 1 x 10 TU/mL

293LTV Lentiviral Cell Line Our 293LTV cell line was selected from the parental 293T cell line for firmer attachment to culture plates and larger, rounder morphology for greater lentiviral production. Recent Product Citations 1. Rossello, R.A. et al. (2013). Mammalian genes induce partially reprogrammed pluripotent stem cells in non-mammalian vertebrate and invertebrate species. eLife Sci. 2:e00036. 2. Dillahunt, S. et al. (2013). Usage of sphingosine kinase isoforms in mast cells is species and/or cell type determined. J. Immunol. 190:2058-2067. Product Name

Size

Catalog Number

LTV-100

1 x 10 Cells

293LTV Cell Line

www.cellbiolabs.com

info@cellbiolabs.com

VIRAL EXPRESSION

Lentiviral Expression

QuickTiter™ Lentivirus Titer / Quantitation Kits Measuring lentiviral titer is important prior to infection of your target cells, and one of the most published methods is the p24 ELISA. Our traditional p24 ELISA kit provides a quick, convenient way to quantify the concentration of your HIV-1 based lentivirus. One disadvantage of using a traditional p24 ELISA to quantify lentivirus is the overexpression of p24 during lentiviral packaging. Free p24 protein may account for a substantial portion of total p24 in lentiviral supernatant. The traditional p24 ELISA detects both virusassociated p24 and free p24 generated by 293T cells during transient transfection. Our QuickTiter™ Lentivirus Titer Kit minimizes the overestimation of p24 in lentivirus supernatant. Our proprietary technology separates the lentivirus-associated p24 from free p24 protein prior to performing the ELISA. If you need a very quick estimate of your lentiviral concentration, try the QuickTiter™ Lentivirus Quantitation Kit. This kit specifically measures the viral nucleic acid content of purified virus or unpurified viral supernatant. This method is ideal for a quick measurement of viral titer, either before or after purification of your lentivirus.

 More Accurate: Exclusive technology in QuickTiter™ Lentivirus Titer Kit minimizes overestimation of virus titer  User-Friendly: Read results on a standard microplate reader

Assay Principle for the QuickTiter™ Lentivirus Titer Kit. Lentivirus particles are packaged with p24 protein, but additional free p24 protein is present in viral supernatant. A traditional p24 ELISA detects both sources of p24 which overestimates viral titer. The QuickTiter™ Lentivirus Titer Kit uses technology to pull the virus out of solution prior to quantitation for a more accurate viral titer.

Selection Guide for QuickTiter™ Lentivirus Quantitation & Titer Kits QuickTiter™ QuickTiter™ Lentivirus Titer Kit Lentivirus Quantitation Kits (Lentivirus-Associated p24 ELISA) (Traditional p24 ELISA) Assay Principle

p24 ELISA with proprietary technology to separate free p24 from viral p24

p24 ELISA

Measures nucleic acid content

Suitable Viruses

Recombinant HIV-1

Recombinant or native HIV-1

HIV-1, FIV, SIV

Detection Method

Colorimetric (ELISA) plate reader

Fluorescence plate reader

Accuracy

Most Published

Speed (45-60 min.)

Key Benefit

QuickTiter™ Lentivirus Quantitation Kit

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Lentiviral Expression

VIRAL EXPRESSION

QuickTiter™ Lentivirus Titer / Quantitation Kits, continued 1.8 p24 in supernatant

1.6

p24 in pellet

OD 450nm

1.4 1.2 1 0.8 0.6 0.4 0.2 0 0

100

p24 (ng/mL) Free p24 Does not Complex with ViraBind™ Reagents. Recombinant p24 was diluted in culture medium and treated with ViraBind™ Lentivirus Reagents A and B found in the QuickTiter™ Lentivirus Titer Kit. The amount of p24 in the supernatant and the pellet was measured according to the assay protocol.

1.2

OD 450 nm

1 0.8 0.6 0.4 0.2 0 Culture medium

GFP Lentiviral Supernatant

p24 Titer of GFP Lentiviral Supernatant. GFP lentiviral construct was cotransfected with a packaging mix into 293 cells. The conditioned medium was harvested 48 hrs after transfection and used to further infect 293 cells. The p24 level of the diluted lentiviral supernatant (1:10 dilution) was determined as described in the assay protocol.

Product Name

Recent Product Citations 1. Belaner, K. et al. (2013). Binding of RNA by APOBEC3G controls deamination-independent restriction of retroviruses. J. Exp. Biol. 216:2213-2220. (VPK-107) 2. Yu, X. et al. (2012). Identification of Hepatitis B virus inhibitors targeting different aspects of infection using a cell-based assay. Antimicrob. Agents Chemother. 56:6109-6120. (VPK-107) 3. Walker, K. et al. (2012). Depletion of GGA1 and GGA3 mediates postinjury elevation of BACE1. J. Neurosci. 32:10423-10437. (VPK-107) 4. Zhou, B. et al. (2012). Interactions between ß-catenin and Transforming growth factor-ß signaling pathways mediate epithelial-mesenchymal transition and are dependent on the transcriptional co-activator cAMP-response element-binding protein (CREB)-binding. J. Biol. Chem. 287:7026-7038. (VPK107) 5. Nedelec, A.D. et al. (2012). Noonan Syndrome-causing SHP2 mutants inhibit insulin-like growth factor 1 release via growth hormone-induced ERK hyperactivation, which contributes to short stature. PNAS 109:4257-4262. (VPK-107) 6. Lavender, H. et al. (2012). In vitro characterization of the activity of PF-05095808, a novel biological agent for Hepatitis C virus therapy. Antimicrob. Agents Chemother. 56:1364-1375. (VPK107) 7. Keck, Z.Y. et al. (2011). Mapping a region of Hepatitis C virus E2 that is responsible for escape from neutralizing antibodies and a core CD81-binding region that does not tolerate neutralization escape mutations. J. Virol. 85:10451-10463. (VPK-107) 8. Sanchez-Antequera, Y. et al. (2011). Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells. Blood 117:e171-e181. (VPK-107) 9. Yi, S.H. et al. (2014). Foxa2 acts as a co-activator potentiating expression of the Nurr1-induced DA phenotype via epigenetic regulation. Development 141:761-772. (VPK-108-H) 10. Smith, B. et al. (2013). Targeting the PyMT oncogene to diverse mammary cell populations enhances tumor heterogeneity and generates rare breast cancer subtypes. Genes & Cancer 10.1177/1947601913475359. (VPK-108-H) 11. Iftikhar, M. et al. (2011). Lysyl oxidase-like-2 (LOXL2) is a major isoform in chondrocytes and is critically required for differentiation. J. Biol. Chem. 286:909-918. (VPK-108-H) 12. Agrawal-Gamse, C. et al. (2010). Yeast-elicited cross-reactive to HIV Env glycans efficiently neutralize virions expressing exclusively high mannose N-linked glycans. J. Virol. 85(1):470-480. (VPK-108-H) 13. Rossello, R.A. et al. (2013). Mammalina genes induce partially reprogrammed pluripotent stem cells in non-mammalian vertebrate and invertebrate species. eLife Sci. 2:e00036. (VPK-112) 14. Fan, X. et al. (2012). Transient, inducible, placenta-specific gene expression in mice. Endocrinology 153:5637-5644. (VPK-112) 15. Veeraraghavalu, K. et al. (2010). Presinilin 1 mutants impair the self-renewal and differentiation of adult murine subventricular zone-neuronal progenitors via cell-autonomous mechanisms involving notch signaling. J. Neurosci. 30:6903-6915. (VPK-112)

Detection

QuickTiter™ Lentivirus Titer Kit (Lentivirus-Associated HIV p24 ELISA)

Colorimetric

QuickTiter™ Lentivirus Quantitation Kit (HIV-1 p24 ELISA)

Colorimetric

QuickTiter™ Lentivirus Quantitation Kit

Fluorometric

www.cellbiolabs.com

Size

Catalog Number

96 Assays

VPK-107

5 x 96 Assays

VPK-107-5

96 Assays

VPK-108-H

5 x 96 Assays

VPK-108-H-5

20 Assays

VPK-112

info@cellbiolabs.com

VIRAL EXPRESSION

Lentiviral Expression

ViraBind™ Lentivirus Concentration & Purification Kits Ultracentrifugation methods used for lentiviral supernatants are tedious and time-consuming and usually only partially purify your virus. Alternatively, filterbased methods have low sample capacity and do not substantially concentrate the virus. ViraBind™ Lentivirus Concentration and Purification Kits produce purified lentivirus with extremely high titer without the need for ultracentrifugation. The virus is pelleted from solution using our proprietary isolation technology, then resuspended in a smaller volume. The resuspended lentivirus is then applied to either a purification column or a dialysis device for purification. If desired, the purified virus may be further concentrated using a centrifugal concentrator.

 Fast: Obtain purified virus in about 4-6 hours with column-based kits and 10-24 hours with dialysis-based kits  High Titer: Concentrate up to 500-fold to as high as 108-1010 TU/ml, sufficient for in vivo studies  High Yield: Recover >60%

Lentivirus Concentration and Purification Procedure.

Selection Guide for Lentivirus Concentration & Purification Kits ViraBind™ Lentivirus Concentration and Purification Kit Purification Method

ViraBind™ PLUS Lentivirus Concentration and Purification Kit

Proprietary Reagent Cocktail Proprietary Reagent Cocktail Proprietary Reagent Cocktail + Purification Column + Dialysis + Dialysis

Total Time Capacity per Prep (Supernatant)

6-8 hours

10-24 hours

100 mL

50 mL

500 mL

Product Name

ViraBind™ Lentivirus Concentration and Purification Kit (100 ml/prep)

ViraBind™ PLUS Lentivirus Concentration and Purification Kit (50 ml/prep) ViraBind™ PLUS Lentivirus Concentration and Purification Mega Kit (500 ml/prep)

ViraBind™ PLUS Lentivirus Concentration and Purification Mega Kit

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

2 Preps

VPK-090

5 Preps

VPK-091

25 Preps

VPK-091-5

2 Preps

VPK-095

2 Preps

VPK-096

10 Preps

VPK-096-5

Fax 1-858-271-6514

VIRAL EXPRESSION

Lentiviral Expression

Recent Product Citations 1. Rossello, R.A. et al. (2013). Mammalina genes induce partially reprogrammed pluripotent stem cells in non-mammalian vertebrate and invertebrate species. eLife Sci. 2:e00036. 2. McEachron, T.A. et al. (2010). Protease-activated receptors mediate crosstalk between coagulation and fibrinolysis. Blood 116:5037-5044. 3. Zemskova, M. et al. (2010). p53-dependent induction of prostate cancer cell senescence by the PIM1 protein kinase. Mol. Cancer Res. 8:1126-1141.

Polybrene

500 400

ViraDuctin™

300 200 100

H T10 80

29 3

H eL a

 Higher Transduction Efficiency: 2-6x higher in many cell lines compared to Polybrene  More Robust: Useful for transduction of nonpermissive cells, including primary cells and stem cells

600

Our ViraDuctin™ Lentivirus Transduction Kit provides superior transduction efficiencies in a variety of cell lines, even when compared to transductions in the presence of Polybrene®.

700

N IH 3T

Lentivirus transduction efficiency is typically low. Additives such as Polybrene® can boost transduction efficiencies, but even then only a small fraction of lentiviral vectors can transduce many target cell lines.

Normalized GFP Fluorescence

ViraDuctin™ Lentivirus Transduction Kit

Transduction Efficiencies in Various Cell Lines. NIH3T3 cells, HeLa cells, our own 293AD cells (page 36) and HT-1080 cells were each seeded at 50,000 cells/well in a 24-well plate overnight. Cells were infected with GFP lentivirus for 48 hours in the presence of Polybrene® or ViraDuctin™ Lentivirus Transduction Kit. For each cell line, fluorescence levels using the ViraDuctin™ system are depicted relative to a normalized fluorescence of 100 for Polybrene®. Polybrene is a registered trademark of Abbott Laboratories.

Transduction of 293AD and HT-1080 Cells. 293AD cells (p. 36) and HT-1080 cells were each seeded at 50,000 cells/well in a 24well plate overnight. Cells were infected with GFP lentivirus for 48 hours in the presence of no additive (left), Polybrene® (middle) or the ViraDuctin™ reagent cocktail (right). Product Name ViraDuctin™ Lentivirus Transduction Kit

Size*

Catalog Number

40 Transductions

LTV-200

200 Transductions

LTV-201

*Based on a 24-well plate. Can also be used with 96-well, 12-well or 6-well plates, as well as 60mm or 100mm dishes. See product insert.

www.cellbiolabs.com

info@cellbiolabs.com

VIRAL EXPRESSION

Retroviral Expression

Retroviral Expression Kits & Reagents Traditional retroviral vectors based on MMLV are useful for integrating genetic material into the host cell genome. However, retrovirus titer tends to be significantly lower than that of adenovirus, which can lead to a lower infection efficiency. Our retroviral reagents and kits incorporate technologies that increase your chances of successful retroviral expression. We offer a comprehensive solution from start to finish:  Retroviral Expression Systems  Gene-Specific Retroviral Vectors  Retroviral Packaging Cell Lines  Concentration / Purification Kits  Retroviral Cloning & Expression  Quantitation / Titer Kits Vectors  Transduction Reagents

Platinum Retroviral Packaging Cell Lines Generate high titers of recombinant retrovirus with a single plasmid transfection* using these extremely powerful, stable cell lines. Platinum Retroviral Packaging Cells are based on the 293T cell line and exhibit greater stability and produce higher yields of retroviral structure proteins, resulting in higher retroviral titers. The Platinum cell lines were invented in the laboratory of Dr. Toshio Kitamura at the University of Tokyo and are available exclusively from Cell Biolabs. They were first described in the following paper: Morita, S. et al. (2000). Gene Therapy 7:1063-1066. Recent Product Citations 1. Schmidt, T. et al. (2013). CXCR4 promotes B cell egress from Peyer’s patches. J. Exp. Med. 10.1084/jem.20122574. (RV-101) 2. Wahlestedt, M. et al. (2013). An epigenetic component of hematopoietic stem cell aging amenable to reprogramming into a young state. Blood 121:4257-4264. (RV-101) 3. Zhong, S. et al. (2013). T-cell receptor affinity and avidity defines antitumor response and autoimmunity in T-cell immunotherapy. PNAS 110:6973-6978. (RV-101) 4. Nam, Y.J. et al. (2013). Reprogramming of human fibroblasts toward a cardiac fate. PNAS 110:5588-5593. (RV-102) 5. Hrdlickova, R. et al. (2012). Alternatively spliced telomerase reverse transcriptase variants lacking telomerase activity stimulate cell proliferation. Mol. Cell Biol. 32:4283-4296. (RV-102) 6. Nowakowski, T. et al. (2013). MicroRNA-92b regulates the development of intermediate cortical progenitors in embryonic mouse brain. PNAS 110:7056-7061. (RV-103) 7. Cavnar, P.J. et al. (2012). The actin regulatory protein HS1 interacts with Arp2/3 and mediates efficient neutrophil chemotaxis. J. Biol. Chem. 287:25466-25477. (RV-103)

Not sure which Platinum Expression System is right for you? See the table below for a selection guide based on the host species of your target cell. Plat-A Cells (Amphotropic)

Plat-E Cells (Ecotropic)

Plat-GP Cells (Pantropic*)

Human

+++

N.S.

+++

Mouse

+++

Rat

+++

Monkey

+++

N.S.

+++

Cat

+++

N.S.

+++

Dog

+++

N.S.

+++

N.S.

+++

Bird

N.S.

+++

Fish

N.S.

+++

Hamster

Frog

N.S.

+++

Insect

N.S.

+++

Mollusk

N.S.

+++

*Plat-GP cells must be co-transfected with a pantropic envelope protein such as VSV-G. N.S. = Not Suitable Suitability of Platinum Retroviral Packaging Cell Lines by Host Species.

Product Name

Size

Platinum-E Retroviral Packaging Cell Line, Ecotropic

>3 x 10 cells

RV-101

Platinum-A Retroviral Packaging Cell Line, Amphotropic

>3 x 106 cells

RV-102

Platinum-GP Retroviral Packaging Cell Line, Pantropic pVSV-G Packaging Vector

Catalog Number

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

>3 x 10 cells

RV-103

10 µg

RV-110

Fax 1-858-271-6514

Retroviral Expression

VIRAL EXPRESSION

Platinum Retroviral Packaging Cells and Expression Systems Our Platinum Retroviral Expression Systems incorporate superior packaging cell lines and vector technologies to produce high-titer virus with a single plasmid transfection. Each Platinum Expression System includes one of our exclusive Platinum Packaging Cell Lines which stably express the gag and pol genes. In the Ecotropic and Amphotropic systems, the packaging cells also express the envelope protein.* Simply clone your gene of interest into the vector provided and transfect into the Platinum cells. Platinum Retroviral Expression Systems contain everything you need to generate your recombinant retrovirus: packaging cell line, expression vector, and GFP control vector. Our pantropic systems also contain a VSVG envelope vector.  Higher Viral Yields: Average titer 107 infectious units/mL with transient transfection  Longer Stability: Expression up to 4 months in the presence of drug selection  Optimized Systems: 3 packaging cell lines for infection of various species; 3 vector backbones (two specifically for infection of stem cells)  Flexible: Order complete systems or cells and vectors separately *Pantropic systems require co-transfection with the provided VSVG envelope vector. Recent Product Citations 1. Aoi, N. et al. (2012). 1a,25-dihydroxyvitamin D3 modulates the hair -inductive capacity of dermal papilla cells: therapeutic potential for hair regeneration. Stem Cells Trans Med. 1:615-626. (VPK-301) 2. Wang, N. et al. (2013). Lacritin rescues stressed epithelia via rapid forkhead box O3 (FOXO3)-associated autophagy that restores metabolism. J. Biol. Chem. 288:18146-18161. (VPK-302) 3. Tanaka, T. et al. (2012). Anthracycline inhibits recruitment of hypoxia-inducible transcription factors and suppresses tumor cell migration and cardiac angiogenic response in the host. J. Biol. Chem. 287:34866-34882. (VPK-302) Product Name

Retrovirus Production Using the Platinum Expression Systems (Ecotropic and Amphotropic).

Expression Vector

Packaging Cell

Catalog Number

Platinum Retroviral Expression System, Ecotropic

pMXs-Puro

Plat-E

VPK-300

Platinum Retroviral Expression System, Amphotropic

pMXs-Puro

Plat-A

VPK-301

Platinum Retroviral Expression System, Pantropic

pMXs-Puro

Plat-GP

VPK-302

Platinum ES/EC Retroviral Expression System, Ecotropic

pMCs-Puro

Plat-E

VPK-303

Platinum ES/EC Retroviral Expression System, Amphotropic

pMCs-Puro

Plat-A

VPK-304

Platinum ES/EC Retroviral Expression System, Pantropic

pMCs-Puro

Plat-GP

VPK-305

Platinum HSC Retroviral Expression System, Ecotropic

pMYs-Puro

Plat-E

VPK-306

Platinum HSC Retroviral Expression System, Amphotropic

pMYs-Puro

Plat-A

VPK-307

Platinum HSC Retroviral Expression System, Pantropic

pMYs-Puro

Plat-GP

VPK-308

www.cellbiolabs.com

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VIRAL EXPRESSION

Retroviral Expression

Retroviral Cloning & Expression Vectors Our Retroviral Expression Vectors are based on backbones derived from Moloney murine leukemia virus (MMLV). We offer the traditional pBABE system and the novel pMXs system, which has been shown to be useful in induced pluripotent stem cell (iPS) studies. pMYs vectors are optimal for use with hematopoietic stem cells, and pMCs vectors are optimal for ES and EC cells. All cloning vectors are supplied as 10 µg in TE buffer.

Retroviral Cloning Vectors for General Gene Expression (driven by 5’ LTR) Recent Product Citations 1. Duran, P.P. et al. (2012). UNG shapes the specificity of AIDinduced somatic hypermutation. J. Exp. Med. 209:1379-1389. (RTV-001-HYGRO) 2. Miyoshi, N. et al. (2010). Defined factors induce reprogramming of gastrointestinal cancer cells. PNAS 107:40-45. (RTV-010) 3. Parikh, C. et al. (2012). Disruption of PH-kinase domain interactions leads to oncogenic activation of AKT in human cancers. PNAS 109:19368-19373. (RTV-012) 4. Zhang, Q. et al. (2013). TNF-a impairs differentiation and function of TGF-ß-induced Treg cells in autoimmune diseases through Akr and Smad3 signaling pathway. J. Mol. Cell Biol. 10.1093/jmcb/jms063. (RTV-013) 5. Sugatani, T. et al. (2011). A microRNA expression signature of osteoclastogenesis. Blood 117:3648-3657. (RTV-014, RTV-016) Vector Name

Cloning Capacity

Catalog Number

pBABEhygro

5.6 kb

RTV-001-HYGRO

pBABEneo

5.9 kb

RTV-003

pBABEpuro

6 kb

RTV-001-PURO

pBABEzeo

6.3 kb

RTV-004

pMXs

5.4 kb

RTV-010

pMXs-IRES-Bsd

5.6 kb

RTV-016

pMXs-IRES-GFP

5.3 kb

RTV-013

pMXs-IRES-Neo

5.2 kb

RTV-015

pMXs-IRES-Puro

5.4 kb

RTV-014

pMXs-Neo

3.8 kb

RTV-011

pMXs-Puro

4.4 kb

RTV-012

pMZs

5.3 kb

RTV-030

Retroviral Cloning Vector for miRNA Recent Product Citation Mansour, M. et al. (2013). The TAL1 complex targets the FBXW7 tumor suppressor by activating miR-223 in human T cell acute lymphoblastic leukemia. J. Exp. Med. 210:1545-1557. (RTV-017) Vector Name pMXs-miR-GFP/Puro

Retroviral Cloning Vectors with Strong Promoters for Overexpression Vector Name

Cloning Capacity

Catalog Number

pMXs-CAG

5.2 kb

RTV-064

pMXs-CMV

5.5 kb

RTV-065

pMXs-EF1

5.5 kb

RTV-063

pMXs-EF1-Bsd

4.2 kb

RTV-062

pMXs-EF1-GFP

3.9 kb

RTV-061

pMXs-EF1-Puro

4 kb

RTV-060

5.4 kb

RTV-066

pMXs-SR

Retroviral Cloning Vectors for use with ES/EC Cells Recent Product Citations 1. Malicet, C. et al. (2011). Distinct properties of human HMGN5 reveal a rapidly evolving but functionally conserved nucleosome binding protein. Mol. Cell Biol. 31:2742-2755. (RTV-040) 2. Mochizunki, Y. et al. (2013). Phosphatidylinositol 3-phosphate myotubularin-related protein 6 (MTMR6) is regulated by small GTPase Rab1b in the early secretory and autophagic pathways. J. Biol. Chem. 288:1009-1021. (RTV-041) Vector Name

Cloning Capacity

Catalog Number

pMCs-IRES-GFP

5.2 kb

RTV-040

pMCs-Puro

4.3 kb

RTV-041

Retroviral Cloning Vectors for use with Hematopoietic Cells Vector Name

Cloning Capacity

Catalog Number

pMYs

5.2 kb

RTV-020

pMYs-IRES-GFP

5.2 kb

RTV-021

pMYs-IRES-Neo

5.2 kb

RTV-023

pMYs-IRES-Puro

5.4 kb

RTV-022

pMYs-Puro

4.3 kb

RTV-024

Retroviral Cloning Vectors for shRNA Vector Name

Cloning Capacity

Catalog Number

pMXs-U6-GFP

5 kb

RTV-071

pMXs-U6-Puro

5.1 kb

RTV-070

Cloning Capacity

Catalog Number

pMXs-U6-Puro-shGFP

RTV-055

4.2 kb

RTV-017

pMXs-U6-Puro-shLuc

RTV-056

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Fax 1-858-271-6514

Retroviral Expression

VIRAL EXPRESSION

Retroviral Packaging Vectors and Cells These constructs are ideal for researchers who prefer a traditional multi-plasmid transfection of 293 cells for packaging recombinant retrovirus. Recent Product Citations 1. Amagai, Y. et al. (2013). Stem cell factor contributes to tumorigenesis of mast cells via an autocrine/paracrine mechanism. J. Leukoc. Biol. 93:245-250. (RV-110) 2. Okamoto, K. et al. (2012). Dengue virus strain DEN2 16681 utilizes a specific glycochain of syndecan-2 proteoglycan as a receptor. J. Gen. Virol. 93:761-770. (RV-110, RV-111) Product Name

Size

Catalog Number

pCMV-10A1 Envelope Vector

100 µL

RV-114

pCMV-Ampho Envelope Vector

100 µL

RV-113

pCMV-Eco Envelope Vector

100 µL

RV-112

pCMV-Gag-Pol Retroviral Vector

10 µg

RV-111

pCMV-VSV-G Envelope Vector

10 µg

RV-110

293RTV Cell Line Our 293RTV cells are derived from the 293 parental cell line, but are selected for firmer attachment to culture plates, faster growth and higher yields of retrovirus produced. Product Name

Size 6

>1 x 10 cells

293RTV Cell Line

Catalog Number RV-100

Gene-Specific Recombinant Retroviral Vectors These constructs are based on backbones derived from MMLV, Vectors with GFP or stem cell factors are supplied as 10 µg of plasmid in TE buffer. All other vectors are supplied as 100 µL of bacterial glycerol stock. Product listing continues on the following pages.

Cell Cycle

Autophagy This vector is supplied with a separate pMXs-GFP control vector at no additional cost. Target Name GFP-LC3

Recent Product Citation Huang, J. et al. (2009). Regulation of the leucocyte chemoattractant receptor FPR in glioblastoma cells by cell differentiation. Carcinogenesis 30(2):348-355. (RTV-401)

Vector Backbone

Catalog Number

pMXs

RTV-801

Reporter Genes

Vector Backbone

Catalog Number

c-Abl

pBABEpuro

RTV-402

c-Abl-TM

pBABEpuro

RTV-403

Recent Product Citations 1. Hrdlickova, R. et al. (2012). Alternatively spliced telomerase reverse transcriptase variants lacking telomerase activity stimulate cell proliferation. Mol. Cell Biol. 32:4283-4296. (RTV-002) 2. Wahlestedt, M. et al. (2013). An epigenetic component of hematopoietic stem cell aging amenable to reprogramming into a young state. Blood 121:4257-4264. (RTV-050)

c-Abl (1-565)

pBABEpuro

RTV-404

Target Name

c-Abl (1-958)

pBABEpuro

RTV-405

pBABEhygro

Target Name

hTERT

p53

Vector Backbone

Catalog Number

GFP

pBABE

RTV-002

RTV-007

GFP

pMCs

RTV-051

pBABEneo

RTV-005

GFP

pMX

RTV-050

pBABEpuro

RTV-006

GFP

pMYs

RTV-052

pBABEpuro

RTV-401

GFP-Puro

pMX

RTV-053

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VIRAL EXPRESSION

Retroviral Expression

Gene-Specific Recombinant Retroviral Vectors, continued MAP Kinase Signaling Vector Name

Vector Backbone

Mutation State

Catalog Number

ERK2

pBABEhygro

Dominant Negative

RTV-109

JNK1

pBABEpuro

Dominant Negative

RTV-110

pBABEpuro

Constitutively Active

RTV-118

pBABEpuro

Dominant Negative

RTV-119

pBABEpuro

Constitutively Active

RTV-120

pBABEpuro

Dominant Negative

RTV-121

pBABEhygro

Constitutively Active

RTV-112

pBABEhygro

Dominant Negative

RTV-111

pBABEpuro

Constitutively Active

RTV-114

pBABEhygro

Dominant Negative

RTV-115

pBABEpuro

Constitutively Active

RTV-116

pBABEhygro

Dominant Negative

RTV-117

pWZLneo

Constitutively Active

RTV-125

p38

pBABEhygro

Dominant Negative

RTV-105

p38

pBABEhygro

Dominant Negative

RTV-106

p38

pBABEhygro

Dominant Negative

RTV-107

p38

pBABEhygro

Dominant Negative

RTV-108

pWZLneo

Constitutively Active

RTV-124

pBABEpuro

Constitutively Active

RTV-122

pBABEpuro

Dominant Negative

RTV-123

pWZLneo

Constitutively Active

RTV-113

MAPKAPK2

MAPKAPK3

MEK1

MKK3

MKK6 myr-Akt1

PI3K p110-CAAX PRAK Raf1-CAAX

Transcription Regulation Target Name

Vector Backbone

Catalog Number

AUF1

pBABEpuro

RTV-305

hnRNPA0

pBABEpuro

RTV-310

hnRNP-A2

pBABEpuro

RTV-340

HuB

pBABEpuro

RTV-302

HuC

pBABEpuro

HuD

Recent Product Citation Yu, Y. et al. (2012). Bcl11a is essential for lymphoid development and negatively regulates p53. J. Exp. Med. 209:2467-2483. (RTV331) Target Name

Vector Backbone

Catalog Number

Stat5A-IRES-GFP

pMXs

RTV-332

RTV-303

Stat5A(1*6)-IRESGFP

pMXs

RTV-333

pBABEpuro

RTV-301

Stat5B

pMXs

RTV-334

HuR

pBABEpuro

RTV-304

Stat5B(1*6)

pMXs

RTV-335

PABP

pBABEpuro

RTV-307

TIA-1

pBABEpuro

RTV-309

Stat5A

pMXs

RTV-330

TIAR

pBABEpuro

RTV-308

Stat5A(1*6)

pMXs

RTV-331

TTP

pBABEpuro

RTV-306

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VIRAL EXPRESSION

Retroviral Expression

Gene-Specific Recombinant Retroviral Vectors, continued Cytoskeleton Regulation Recent Product Citation Zhao, B. et al. (2012). TNF-induced osteoclastogenesis and inflammatory bone resorption are inhibited by transcription factor RBP-J. J. Exp. Med. 209:2467-2483. (RTV-101) Target Name Cdc42

Vector Backbone

Mutation State

Catalog Number

pBABEhygro

L61

RTV-203

pBABEpuro

N/A

RTV-220

pWZLhygro

Q61

RTV-221

N/A

RTV-201

V12

RTV-206

K-Ras

myr-Rac1

pBABEpuro

Rac1

pBABEhygro

V12

RTV-202

N-Ras

pBABEpuro

K61

RTV-222

Rac3

pBABEhygro

V12

RTV-205

pBABEpuro

V12

RTV-101

pBABEpuro

V12C40

RTV-104

pBABEpuro

V12G37

RTV-103

pBABEpuro

V12S35

RTV-102

pBABEhygro

L63

RTV-204

Ras

RhoA

iPS / Stem Cell Factors Human iPS Genes

Mouse iPS Genes

Target Name

Vector Backbone

Catalog Number

Target Name

Vector Backbone

Catalog Number

4-Vector Set*

pMXs

RTV-701-C

4-Vector Set*

pMXs

RTV-705-C

6-Vector Set**

pMXs

RTV-709-C

6-Vector Set**

pMXs

RTV-711-C

c-Myc

pMXs

RTV-703

c-Myc

pMXs

RTV-707

Klf4

pMXs

RTV-704

Klf4

pMXs

RTV-708

Lin-28

pMXs

RTV-710

Lin-28

pMXs

RTV-712

NANOG

pMXs

RTV-709

NANOG

pMXs

RTV-711

Oct-3/4

pMXs

RTV-701

Oct-3/4

pMXs

RTV-705

Sox2

pMXs

RTV-702

Sox2

pMXs

RTV-706

p53 shRNA

pRetro

RTV-410

p53 shRNA

pRetro

RTV-400

*4-Vector sets contain individual constructs with the following genes: c-Myc, Klf4, Oct-3/4 and Sox2. **6-Vector sets contain individual constructs with the following genes: c-Myc, Klf4, Oct-3/4, Sox2, Lin-28 and NANOG.

Proteases and Related Molecules Target Name

Vector Backbone

Catalog Number

uPA

pBABEpuro

RTV-501

uPAR

pBABEhygro

RTV-502

Recent Product Citation Gutova, M. et al (2008). Urokinase plasminogen activator and urokinase plasminogen activator receptor mediate human stem cell tropism to malignant solid tumors. Stem Cells 26:1406-1413. (RTV-501, RTV-502)

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VIRAL EXPRESSION

Retroviral Expression

ViraBind™ Retrovirus Concentration & Purification Kits Ultracentrifugation methods used for lentiviral supernatants are tedious and time-consuming and usually only partially purify your virus. Alternatively, filterbased methods have low sample capacity and do not substantially concentrate the virus. ViraBind™ Retrovirus Concentration and Purification Kits produce purified lentivirus with extremely high titer without the need for ultracentrifugation. The virus is pelleted from solution using our proprietary isolation technology, then resuspended in a smaller volume. The resuspended retrovirus is then applied to either a purification column or a dialysis device for purification. If desired, the purified virus may be further concentrated using a centrifugal concentrator.

 Fast: Obtain purified virus in about 4-6 hours  High Titer: Concentrate 500-fold to 109-1010 TU/ml, sufficient for in vivo studies  High Yield: Recover >60%  High Throughput: Process greater volumes per prep than filter-based purification methods Retrovirus Concentration and Purification Procedure.

Selection Guide for Retrovirus Concentration & Purification Kits ViraBind™ Retrovirus Concentration and Purification Kit Purification Method

ViraBind™ PLUS Retrovirus Concentration and Purification Kit

Proprietary Reagent Cocktail Proprietary Reagent Cocktail Proprietary Reagent Cocktail + Purification Column + Dialysis + Dialysis

Total Time Capacity per Prep (Supernatant)

6-8 hours

10-24 hours

100 mL

50 mL

500 mL

Product Name

ViraBind™ Retrovirus Concentration and Purification Kit (100 ml/prep)

ViraBind™ PLUS Retrovirus Concentration and Purification Kit (50 ml/prep) ViraBind™ PLUS Retrovirus Concentration and Purification Mega Kit (500 ml/prep)

ViraBind™ PLUS Retrovirus Concentration and Purification Mega Kit

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

2 Preps

VPK-130

5 Preps

VPK-131

25 Preps

VPK-131-5

2 Preps

VPK-135

2 Preps

VPK-136

10 Preps

VPK-136-5

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VIRAL EXPRESSION

Retroviral Expression QuickTiter™ Retrovirus Rapid Quantitation Kit This kit specifically measures the viral nucleic acid content of purified virus or unpurified viral supernatant. This method is ideal for a quick measurement of viral titer, either before or after purification of your retrovirus.  Ultra-fast Results: 45-60 minute procedure  Convenient: Titer may be measured before purification step  Sensitive: Limit of detection = 1.5 x 109 VP/mL from 2 mL of retroviral supernatant 500

RFU (520 nm)

400 300 200 100 0 0

400

800

1200

Retroviral RNA (ng)

RFU (520 nm)

70 60 50 40 30 20 10 0 0

100

150

Retroviral RNA (ng) Retrovirus RNA Standard Curve. The QuickTiter™ Retrovirus RNA Standard was diluted according to the assay protocol. Fluorescence was measured on a SpectraMax Gemini XS Fluorometer (Molecular Devices) with a 485 / 538 nm filter set and a 530 nm cutoff. Recent Product Citation Ito, T. et al. (2012). Stem cell factor programs the mast cell activation phenotype. J. Immunol. 188:5428-5437. Product Name QuickTiter™ Retrovirus Quantitation Kit

Assay Procedure for the QuickTiter™ Retrovirus Quantitation Kit. Detection

Size

Catalog Number

Fluorometric

20 Assays

VPK-120

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VIRAL EXPRESSION

Retroviral Expression

ViraDuctin™ Retrovirus Transduction Kit The efficiency of retrovirus transduction can be low compared to other viruses. The rate at which retroviral vectors bind to cells is controlled mostly by diffusion. Additionally, the presence of transduction inhibitors such as proteoglycans and glycosaminoglycans in retroviral supernatants can lead to poor gene transfer. Additives such as Polybrene® can boost transduction efficiencies, but they do not eliminate these transduction inhibitors.

 More Robust: Removes harmful transduction inhibitors from retroviral supernatant  Higher Transduction Efficiencies: Compared to infections in the presence of Polybrene or no additives  Versatile: Particularly useful for nonpermissive cells including primary cells and stem cells, but may boost transduction rates in a wide variety of cells

Our ViraDuctin™ Retrovirus Transduction Kit provides superior transduction efficiencies even when compared to transductions in the presence of Polybrene®. A proprietary reagent cocktail forms a supercomplex with the retrovirus which is pelleted away from the supernatant, removing detrimental transduction inhibitors that decrease infection efficiency.

Recent Product Citations 1. Gandhi, M. et al. (2012). Homologous chromosomes make contact at the sites of double-strand breaks in genes in somatic G0/ G1-phase human cells. PNAS 109:9454-9459. 2. Miyoshi, N. et al. (2010). Defined factors induce reprogramming of gastrointestinal cancer cells. PNAS 107:40-45.

Product Name ViraDuctin™ Retrovirus Transduction Kit

Size*

Catalog Number

40 Transductions

RV-200

200 Transductions

RV-201

*Number of transductions shown is based on use in a 24-well plate. This product may also be used with 96-well, 12-well or 6-well plates, as well as 60mm or 100mm dishes. See product insert for specific details.

Polybrene is a registered trademark of Abbott Laboratories.

Phone 1-858-271-6500

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MICRORNA

ANALYSIS

miRNA Clone Collection

miRNASelect™ Precursor Clone Collection (Expression Vectors) miRNASelect™ Expression Vectors contain full miRNA precursor sequences with constitutive promoter.  Human (hsa) vectors contain a puromycin selection marker  Mouse (mmu) vectors contain a GFP-puromycin fusion

Don’t see your microRNA of interest? Clone your own sequence into one of our Expression Vectors; see page 80. Recent Product Citations 1. Dar, A. et al. (2013). The role of miR-18b in MDM2-p53 pathway signaling and melanoma progression. J. Natl. Cancer Inst. 105:433-442. (MIR-18B) 2. Mallory, M.J. et al. (2011). Signal– and developmental-dependent alternative splicing of LEF1 in T cells is controlled by CELF2. Mol. Cell. Biol. 31:2184-2195. (MIR-23B) 3. Woo, H. et al. (2013). Nucleolin mediates microRNA-directed CSF-1 mRNA deadenylation but increases translation of CSF-1 mRNA. Mol. Cell Proteomics 12:16611667. (MIR-130A, MIR-301A) 4. Starega-Roslan, J. et al. (2010). Stuctural basis of microRNA length variety. Nuc. Acids Res. 10.1093/nar/gkq727. (MIR148A) 5. Saus, E. et al. (2010). Genetic variants and abnormal processing of pre-miR-182, a circadian clock modulator, in major depression patients with late insomnia. Hum. Mol. Genet. 19:4017. (MIR-182) 6. Beezhold, K. et al. (2011). miR-190mediated downregulation of PHLPP contributes to arsenic-induced Akt activation and carcinogenesis. Toxicol. Sci. 123:411420. (MIR-190) 7. Wang, Y.S. et al. (2012). MicroRNA-195 regulates vascular smooth muscle cell phenotype and prevents neointimal formation. Cardiovasc. Res. 95:5174-526. (MIR195) 8. Majid, S. et al. (2011). MicroRNA-205 inhibits src-mediated oncogenic pathways in renal cancer. Cancer Res. 71:26112621. (MIR-205) 9. Halappanaver, S. et al. (2013). IL-1 receptor regulates microRNA-135b expression in a negative feedback mechanism. J. Immunol. 190:3679-3686. (MMU-MIR135B) 10.Yamamoto, H. et al. (2012). MicroRNA494 regulates mitochondrial biogenesis in skeletal muscle through mitochondrial transcription factor A and forkhead box j3. Am J. Physiol. Endocrinol. Metab. 303:E1419-1427. (MMU-MIR-494)

Phone 1-858-271-6500

Precursor Name

Catalog Number

Precursor Name

Catalog Number

mmu-let-7a-1

MMU-LET7A-1

hsa-mir-16-1

hsa-let-7a-2

MIR-LET7A-2

mmu-mir-16-1

hsa-let-7a-3

MIR-LET7A-3

hsa-mir-16-2

mmu-let-7b

MMU-LET7B

hsa-mir-17

MIR-17

hsa-let7c

MIR-LET7C

hsa-mir-18a

MIR-18A

mmu-let-7c-1

MMU-LET7C-1

mmu-mir-18a

mmu-let-7c-2

MMU-LET7C-2

hsa-mir-18b

hsa-let-7d

MIR-LET7D

mmu-mir-18b

mmu-let-7d

MMU-LET7D

hsa-mir-19a

hsa-let-7e

MIR-16-1 MMU-MIR-16-1 MIR-16-2

MMU-MIR-18A MIR-18B MMU-MIR-18B MIR-19A

MIR-LET7E

mmu-mir-19a

MMU-MIR-19A

hsa-let-7f-1

MIR-LET7F-1

hsa-mir-19b-1

MIR-19B-1

mmu-let-7f-1

MMU-LET7F-1

mmu-mir-19b-1

mmu-let-7f-2

MMU-LET7F-2

hsa-mir-19b-2

hsa-let-7g

MIR-LET7G

mmu-mir-19b-2

mmu-let-7g

MMU-LET7G

hsa-mir-20a

hsa-let-7i hsa-mir-1-1 mmu-mir-1-1 hsa-mir-1-2 mmu-mir-1-2

MIR-LET7I MIR-1-1 MMU-MIR-1-1 MIR-1-2 MMU-MIR-1-2

mmu-mir-20a hsa-mir-20b mmu-mir-20b hsa-mir-21

MMU-MIR-19B-1 MIR-19B-2 MMU-MIR-19B-2 MIR-20A MMU-MIR-20A MIR-20B MMU-MIR-20B MIR-21

mmu-mir-21

MMU-MIR-21

hsa-mir-7-1

MIR-7-1

hsa-mir-22

MIR-22

hsa-mir-7-2

MIR-7-2

hsa-mir-23b

MIR-23B

hsa-mir-7-3

MIR-7-3

mmu-mir-23b

MMU-MIR-23B

mmu-mir-7a-1

MMU-MIR-7A-1

mmu-mir-24-1

MMU-MIR-24-1

mmu-mir-7a-2

MMU-MIR-7A-2

hsa-mir-24-2

mmu-mir-7b

MMU-MIR-7B

hsa-mir-9-1

MIR-9-1

mmu-mir-9-1

MMU-MIR-9-1

mmu-mir-24-2 mmu-mir-25 hsa-mir-26a-1

hsa-mir-9-2

MIR-9-2

mmu-mir-26a-1

hsa-mir-10a

MIR-10A

hsa-mir-26a-2

hsa-mir-10b

MIR-10B

mmu-mir-26a-2

mmu-mir-10b

MIR-24-2 MMU-MIR-24-2 MMU-MIR-25 MIR-26A-1 MMU-MIR-26A-1 MIR-26A-2 MMU-MIR-26A-2

MMU-MIR-10B

hsa-mir-26b

MIR-26B

MIR-15A

hsa-mir-27a

MIR-27A

mmu-mir-15a

MMU-MIR-15A

hsa-mir-27b

MIR-27B

mmu-mir-15b

MMU-MIR-15B

mmu-mir-27b

hsa-mir-15a

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MMU-MIR-27B

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miRNA Clone Collection

MICRORNA

ANALYSIS

miRNASelectâ&#x201E;˘ Precursor Clone Collection (Expression Vectors), cont. Precursor Name hsa-mir-28

Catalog Number MIR-28

Precursor Name mmu-mir-96 hsa-mir-98

Catalog Number MMU-MIR-96 MIR-98

Precursor Name hsa-mir-129-2

mmu-mir-28

MMU-MIR-28

hsa-mir-29a

MIR-29A

mmu-mir-98

MMU-MIR-98

mmu-mir-29a

MMU-MIR-29A

hsa-mir-100

MIR-100

hsa-mir-29b-1

MIR-29B-1

mmu-mir-100

MMU-MIR-100

mmu-mir-29b-1

MMU-MIR-29B-1

mmu-mir-101a

MMU-MIR-101A

mmu-mir-130b

mmu-mir-29b-2

MMU-MIR-29B-2

mmu-mir-101b

MMU-MIR-101B

hsa-mir-132

MIR-29C

hsa-mir-103-1

MIR-103-1

hsa-mir-29c

mmu-mir-30c-1 hsa-mir-30c-2 mmu-mir-30c-2 hsa-mir-30d mmu-mir-30d hsa-mir-30e mmu-mir-30e hsa-mir-31 mmu-mir-31

mmu-mir-132

MIR-130B MMU-MIR-130B MIR-132 MMU-MIR-132

MIR-103-2

mmu-mir-133a-2

MMU-MIR-133A-2

MMU-MIR-30A

mmu-mir-105

MMU-MIR-105

mmu-mir-133b

MMU-MIR-133B

MIR-30B

hsa-mir-105-1

MIR-105-1

mmu-mir-134

MMU-MIR-134

MIR-30C-1

hsa-mir-105-2

MIR-105-2

hsa-mir-135a-1

MMU-MIR-30C-1

hsa-mir-106a

MIR-106A

mmu-mir-135a-1

MIR-30C-2 MMU-MIR-30C-2 MIR-30D MMU-MIR-30D

mmu-mir-106a hsa-mir-106b mmu-mir-106b hsa-mir-107

MMU-MIR-106A MIR-106B MMU-MIR-106B MIR-107

hsa-mir-135a-2 mmu-mir-135a-2 hsa-mir-135b mmu-mir-135b

MIR-30E

mmu-mir-107

MMU-MIR-107

hsa-mir-136

MMU-MIR-30E

mmu-mir-122

MMU-MIR-122

mmu-mir-136

MIR-31 MMU-MIR-31

mmu-mir-124-1 hsa-mir-124-2

MMU-MIR-124-1

hsa-mir-137

MMU-MIR-135A-1 MIR-135A-2 MMU-MIR-135A-2 MIR-135B MMU-MIR-135B MIR-136 MMU-MIR-136 MIR-137

mmu-mir-137

MMU-MIR-137 MIR-138-1

mmu-mir-124-2

MMU-MIR-124-2

hsa-mir-138-1

mmu-mir-32

MMU-MIR-32

mmu-mir-124-3

MMU-MIR-124-3

mmu-mir-138-1

hsa-mir-33a

MIR-33A

mmu-mir-125a

MMU-MIR-125A

hsa-mir-138-2

hsa-mir-34c

MIR-34C

hsa-mir-125b-2

MIR-125B-2

mmu-mir-125b-2

MIR-135A-1

MIR-124-2

MIR-32

hsa-mir-32

MMU-MIR-130A

hsa-mir-103-2

MIR-30A

hsa-mir-30c-1

hsa-mir-130b

MIR-130A

MMU-MIR-133A-1

hsa-mir-30a

hsa-mir-30b

mmu-mir-130a

MMU-MIR-129-2

mmu-mir-133a-1

MMU-MIR-29C

mmu-mir-30a

hsa-mir-130a

MIR-129-2

MMU-MIR-103-1

mmu-mir-29c

mmu-mir-103-1

mmu-mir-129-2

Catalog Number

MMU-MIR-125B-2

mmu-mir-138-2

MIR-138-2 MMU-MIR-138-2

mmu-mir-34c

MMU-MIR-34C

hsa-mir-92a-1

MIR-92A-1

mmu-mir-126

MMU-MIR-126

mmu-mir-139

MMU-MIR-92A-1

mmu-mir-127

MMU-MIR-127

hsa-mir-140

MIR-92A-2

hsa-mir-128-1

MIR-128-1

mmu-mir-140

MMU-MIR-140

MMU-MIR-128-1

mmu-mir-141

MMU-MIR-141

mmu-mir-92a-1 hsa-mir-92a-2 mmu-mir-92a-2

MMU-MIR-92A-2

mmu-mir-92b

MMU-MIR-92B

mmu-mir-93

MMU-MIR-93

mmu-mir-128-1 hsa-mir-128-2 mmu-mir-128-2

hsa-mir-95

MIR-95

hsa-mir-129-1

hsa-mir-96

MIR-96

mmu-mir-129-1

MIR-128-2 MMU-MIR-128-2 MIR-129-1 MMU-MIR-129-1

www.cellbiolabs.com

hsa-mir-139

MMU-MIR-138-1

hsa-mir-142 mmu-mir-142 hsa-mir-143 mmu-mir-143

MIR-139 MMU-MIR-139 MIR-140

MIR-142 MMU-MIR-142 MIR-143 MMU-MIR-143

info@cellbiolabs.com

MICRORNA

ANALYSIS

miRNA Clone Collection

miRNASelectâ&#x201E;˘ Precursor Clone Collection (Expression Vectors), cont. Precursor Name mmu-mir-144 hsa-mir-145

MMU-MIR-144 MIR-145

Precursor Name mmu-mir-188 hsa-mir-190

Catalog Number

Precursor Name

Catalog Number

MMU-MIR-188

mmu-mir-206

MMU-MIR-206

MIR-190

hsa-mir-208a

MIR-208A

mmu-mir-145

MMU-MIR-145

mmu-mir-190

MMU-MIR-190

mmu-mir-208a

MMU-MIR-208A

mmu-mir-146a

MMU-MIR-146A

mmu-mir-190b

MMU-MIR-190B

mmu-mir-208b

MMU-MIR-208B

hsa-mir-146b

MIR-146B

mmu-mir-191

MMU-MIR-191

mmu-mir-210

MMU-MIR-210

hsa-mir-147

MIR-147

mmu-mir-192

MMU-MIR-192

hsa-mir-211

hsa-mir-147b

MIR-147B

mmu-mir-193

MMU-MIR-193

mmu-mir-211

hsa-mir-148a

MIR-148A

mmu-mir-193b

MMU-MIR-193B

mmu-mir-148a

MMU-MIR-148A

hsa-mir-194-1

MIR-194-1

mmu-mir-149

MMU-MIR-149

mmu-mir-194-1

MMU-MIR-194-1

hsa-mir-214

MIR-150

mmu-mir-194-2

MMU-MIR-194-2

mmu-mir-214

MMU-MIR-214

MIR-195

mmu-mir-215

MMU-MIR-215

MIR-196A-1

hsa-mir-216a

MIR-216A

hsa-mir-150 mmu-mir-150 hsa-mir-151 mmu-mir-151 hsa-mir-152

MMU-MIR-150 MIR-151 MMU-MIR-151 MIR-152

hsa-mir-195 hsa-mir-196a-1 mmu-mir-196a-1 hsa-mir-196a-2

mmu-mir-152

MMU-MIR-152

mmu-mir-196a-2

mmu-mir-153

MMU-MIR-153

mmu-mir-196b

hsa-mir-154

hsa-mir-212 mmu-mir-212

MMU-MIR-216B

MMU-MIR-196A-2

mmu-mir-217

MMU-MIR-217

MMU-MIR-196B

mmu-mir-218-1

MMU-MIR-218-1 MMU-MIR-218-2

MMU-MIR-154

hsa-mir-198

MIR-198

hsa-mir-219-1

mmu-mir-155

MMU-MIR-155

hsa-mir-199a-1

hsa-mir-199a-2

mmu-mir-181a-2

MMU-MIR-181A-2

hsa-mir-200a

mmu-mir-181b-1

MMU-MIR-181B-1

mmu-mir-181b-2

MMU-MIR-181B-2

mmu-mir-181c

MMU-MIR-181C

hsa-mir-181d

MIR-181D

mmu-mir-182

MIR-214

mmu-mir-216b

mmu-mir-154

MMU-MIR-181A-1

MMU-MIR-212

MIR-196A-2

mmu-mir-218-2

mmu-mir-181a-1

MIR-212

MMU-MIR-216A

MIR-197

mmu-mir-199a-1

MMU-MIR-211

mmu-mir-216a

hsa-mir-197

MIR-181A-1

MIR-211

MMU-MIR-196A-1

MIR-154

hsa-mir-181a-1

MIR-219-1

MIR-199A-1

mmu-mir-219-1

MMU-MIR-219-1

MMU-MIR-199A-1

mmu-mir-219-2

MMU-MIR-219-2

MIR-199A-2

hsa-mir-222

MIR-222

MIR-200A

mmu-mir-222

MMU-MIR-222

mmu-mir-200a

MMU-MIR-200A

mmu-mir-223

MMU-MIR-223

mmu-mir-200b

MMU-MIR-200B

hsa-mir-224 mmu-mir-224

MMU-MIR-224

mmu-mir-200c

MMU-MIR-200C

mmu-mir-290

MMU-MIR-290

MMU-MIR-182

mmu-mir-201

MMU-MIR-201

mmu-mir-291a

MMU-MIR-291A

mmu-mir-183

MMU-MIR-183

hsa-mir-202

MIR-202

mmu-mir-291b

MMU-MIR-291B

mmu-mir-184

MMU-MIR-184

mmu-mir-202

MMU-MIR-202

mmu-mir-292

MMU-MIR-292

MIR-204

mmu-mir-293

MMU-MIR-293

MMU-MIR-204

mmu-mir-294

MMU-MIR-294

MIR-205

mmu-mir-295

MMU-MIR-295

MMU-MIR-205

mmu-mir-296

MMU-MIR-296

mmu-mir-185

MIR-185 MMU-MIR-185

hsa-mir-200c

MIR-224

MIR-200C

hsa-mir-185

hsa-mir-204 mmu-mir-204

hsa-mir-186

MIR-186

hsa-mir-205

hsa-mir-187

MIR-187

mmu-mir-205

mmu-mir-187

Catalog Number

MMU-MIR-187

Phone 1-858-271-6500

hsa-mir-206

MIR-206

USA Toll-Free 1-888-CBL-0505

hsa-mir-297

MIR-297

Fax 1-858-271-6514

miRNA Clone Collection

MICRORNA

ANALYSIS

miRNASelectâ&#x201E;˘ Precursor Clone Collection (Expression Vectors), cont. Precursor Name

Catalog Number

Precursor Name

mmu-mir-297a-1

MMU-MIR-297A-1

mmu-mir-343

mmu-mir-297a-3

MMU-MIR-297A-3

mmu-mir-297a-4

Catalog Number

Precursor Name

Catalog Number

MMU-MIR-343

mmu-mir-433

MMU-MIR-433

mmu-mir-344-1

MMU-MIR-344-1

mmu-mir-434

MMU-MIR-434

MMU-MIR-297A-4

mmu-mir-344-2

MMU-MIR-344-2

mmu-mir-448

MMU-MIR-448

mmu-mir-297a-5

MMU-MIR-297A-5

mmu-mir-345

MMU-MIR-345

mmu-mir-449a

MMU-MIR-449A

mmu-mir-297a-6

MMU-MIR-297A-6

mmu-mir-346

MMU-MIR-346

mmu-mir-449b

MMU-MIR-449B

mmu-mir-297b

MMU-MIR-297B

mmu-mir-351

MMU-MIR-351

mmu-mir-449c

MMU-MIR-449C

mmu-mir-297c

MMU-MIR-297C

mmu-mir-361

MMU-MIR-361

mmu-mir-450a-2

MIR-298

mmu-mir-363

MMU-MIR-363

hsa-mir-450b

MIR-450B

hsa-mir-298

MMU-MIR-450A-2

mmu-mir-298

MMU-MIR-298

mmu-mir-365-1

MMU-MIR-365-1

mmu-mir-451

MMU-MIR-451

mmu-mir-299

MMU-MIR-299

mmu-mir-365-2

MMU-MIR-365-2

mmu-mir-452

MMU-MIR-452

hsa-mir-301a

MIR-301A

mmu-mir-367

MMU-MIR-367

mmu-mir-455

MMU-MIR-455

hsa-mir-301b

MIR-301B

mmu-mir-370

MMU-MIR-370

mmu-mir-464

MMU-MIR-464

hsa-mir-302a

MIR-302A

mmu-mir-374

MMU-MIR-374

mmu-mir-465a

MMU-MIR-465A

MMU-MIR-302A

hsa-mir-374b

MIR-374B

mmu-mir-465b-1

MMU-MIR-465B-1

MIR-302B

mmu-mir-375

MMU-MIR-375

mmu-mir-465b-2

MMU-MIR-465B-2

MMU-MIR-302B

mmu-mir-376a

MMU-MIR-376A

mmu-mir-465c-1

MMU-MIR-465C-1

MIR-302C

mmu-mir-376c

MMU-MIR-376C

mmu-mir-465c-2

MMU-MIR-465C-2

mmu-mir-302c

MMU-MIR-302C

mmu-mir-377

MMU-MIR-377

mmu-mir-466a

mmu-mir-302d

MMU-MIR-302D

mmu-mir-378

MMU-MIR-378

mmu-mir-466b-2

MMU-MIR-466B-2

mmu-mir-320

MMU-MIR-320

mmu-mir-379

MMU-MIR-379

mmu-mir-466b-3

MMU-MIR-466B-3

mmu-mir-322

MMU-MIR-322

mmu-mir-380

MMU-MIR-380

mmu-mir-466d

MIR-323

mmu-mir-381

MMU-MIR-381

mmu-mir-466f-1

MMU-MIR-466F-1

mmu-mir-324

MMU-MIR-324

mmu-mir-382

MMU-MIR-382

mmu-mir-466f-4

MMU-MIR-466F-4

mmu-mir-325

MMU-MIR-325

mmu-mir-383

MMU-MIR-383

mmu-mir-466g

MMU-MIR-466G

mmu-mir-328

MMU-MIR-328

mmu-mir-384

MMU-MIR-384

mmu-mir-466h

MMU-MIR-466H

hsa-mir-329-1

MIR-329-1

mmu-mir-409

MMU-MIR-409

mmu-mir-466i

MMU-MIR-466I

mmu-mir-330

MMU-MIR-330

mmu-mir-410

MMU-MIR-410

mmu-mir-466j

MMU-MIR-466J

MIR-331

mmu-mir-411

MMU-MIR-411

mmu-mir-466k

MMU-MIR-466K

MMU-MIR-331

mmu-mir-412

MMU-MIR-412

mmu-mir-466l

MMU-MIR-466L

MIR-335

mmu-mir-421

MMU-MIR-421

mmu-mir-467b

MMU-MIR-467B

mmu-mir-337

MMU-MIR-337

mmu-mir-423

MMU-MIR-423

mmu-mir-467d

MMU-MIR-467D

mmu-mir-338

MMU-MIR-338

mmu-mir-425

MMU-MIR-425

mmu-mir-467e

MMU-MIR-467E

mmu-mir-341

MMU-MIR-341

mmu-mir-429

MMU-MIR-429

mmu-mir-467f

MMU-MIR-467F

MIR-342

mmu-mir-431

MMU-MIR-431

mmu-mir-467g

MMU-MIR-467G

MIR-433

mmu-mir-469

MMU-MIR-469

mmu-mir-302a hsa-mir-302b mmu-mir-302b hsa-mir-302c

hsa-mir-323

hsa-mir-331 mmu-mir-331 hsa-mir-335

hsa-mir-342 mmu-mir-342

MMU-MIR-342

hsa-mir-433

www.cellbiolabs.com

MMU-MIR-466A

MMU-MIR-466D

info@cellbiolabs.com

MICRORNA

ANALYSIS

miRNA Clone Collection

miRNASelectâ&#x201E;˘ Precursor Clone Collection (Expression Vectors), cont. Precursor Name

Precursor Name

mmu-mir-483

MMU-MIR-483

mmu-mir-541

mmu-mir-484

MMU-MIR-484

hsa-mir-542

mmu-mir-485

MMU-MIR-485

mmu-mir-542

mmu-mir-486

MMU-MIR-486

hsa-mir-543

mmu-mir-487b

MMU-MIR-487B

mmu-mir-488

Catalog Number

Product Name

Catalog Number

MMU-MIR-541

hsa-mir-600

MIR-600

MIR-542

hsa-mir-601

MIR-601

MMU-MIR-542

hsa-mir-602

MIR-602

MIR-543

hsa-mir-603

MIR-603

mmu-mir-546

MMU-MIR-546

hsa-mir-605

MIR-605

MMU-MIR-488

mmu-mir-547

MMU-MIR-547

hsa-mir-606

MIR-606

mmu-mir-489

MMU-MIR-489

hsa-mir-548a-1

MIR-548A-1

hsa-mir-608

MIR-608

mmu-mir-490

MMU-MIR-490

hsa-mir-548a-3

MIR-548A-3

hsa-mir-609

MIR-609

MIR-491

hsa-mir-548b

MIR-548B

hsa-mir-610

MIR-610

mmu-mir-491

MMU-MIR-491

hsa-mir-548c

MIR-548C

hsa-mir-613

MIR-613

mmu-mir-493

MMU-MIR-493

hsa-mir-548d-1

MIR-548D-1

hsa-mir-616

MIR-616

mmu-mir-494

MMU-MIR-494

hsa-mir-548d-2

MIR-548D-2

hsa-mir-619

MIR-619

mmu-mir-495

MMU-MIR-495

hsa-mir-551b

MIR-551B

hsa-mir-620

MIR-620

mmu-mir-496

MMU-MIR-496

hsa-mir-554

MIR-554

hsa-mir-628

MIR-628

mmu-mir-497

MMU-MIR-497

hsa-mir-555

MIR-555

hsa-mir-630

MIR-630

MIR-499

hsa-mir-568

MIR-568

hsa-mir-633

MIR-633

MMU-MIR-568

hsa-mir-635

MIR-635

hsa-mir-491

hsa-mir-499 mmu-mir-499

MMU-MIR-499

mmu-mir-568

mmu-mir-501

MMU-MIR-501

hsa-mir-569

MIR-569

hsa-mir-636

MIR-636

MIR-503

hsa-mir-577

MIR-577

hsa-mir-637

MIR-637

MMU-MIR-503

hsa-mir-579

MIR-579

hsa-mir-641

MIR-641

MIR-504

hsa-mir-580

MIR-580

hsa-mir-643

MIR-643

MMU-MIR-504

hsa-mir-581

MIR-581

hsa-mir-645

MIR-645

MIR-505

hsa-mir-582

MIR-582

hsa-mir-649

MIR-649

MMU-MIR-582

hsa-mir-651

MIR-651 MIR-652

hsa-mir-503 mmu-mir-503 hsa-mir-504 mmu-mir-504 hsa-mir-505 mmu-mir-505

MMU-MIR-505

mmu-mir-582

hsa-mir-506

MIR-506

hsa-mir-584

MIR-584

hsa-mir-652

hsa-mir-508

MIR-508

hsa-mir-585

MIR-585

mmu-mir-652

hsa-mir-509-1

MIR-509-1

hsa-mir-591

MIR-591

hsa-mir-653

hsa-mir-514-1

MIR-514-1

hsa-mir-592

MIR-592

mmu-mir-653

hsa-mir-514-2

MIR-514-2

mmu-mir-592

hsa-mir-520f

MIR-520F

hsa-mir-525

MMU-MIR-652 MIR-653 MMU-MIR-653

MMU-MIR-592

hsa-mir-654

MIR-654

hsa-mir-595

MIR-595

hsa-mir-658

MIR-658

MIR-525

hsa-mir-596

MIR-596

hsa-mir-659

MIR-659

MIR-526A-2

hsa-mir-597

MIR-597

hsa-mir-665

MIR-665

mmu-mir-539

MMU-MIR-539

hsa-mir-598

MIR-598

mmu-mir-665

MMU-MIR-665

mmu-mir-540

MMU-MIR-540

mmu-mir-598

MMU-MIR-598

mmu-mir-666

MMU-MIR-666

MIR-599

mmu-mir-667

MMU-MIR-667

hsa-mir-526a-2

hsa-mir-541

Catalog Number

MIR-541

Phone 1-858-271-6500

hsa-mir-599

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

miRNA Clone Collection

MICRORNA

ANALYSIS

miRNASelectâ&#x201E;˘ Precursor Clone Collection (Expression Vectors), cont. Product Name mmu-mir-668 mmu-mir-669a-3

Catalog Number MMU-MIR-668 MMU-MIR-669A-3

Product Name mmu-mir-705 hsa-mir-708

Catalog Number MMU-MIR-705 MIR-708

Product Name hsa-mir-876 mmu-mir-876

Catalog Number MIR-876 MMU-MIR-876

mmu-mir-669b

MMU-MIR-669B

mmu-mir-708

MMU-MIR-708

hsa-mir-877

mmu-mir-669c

MMU-MIR-669C

mmu-mir-711

MMU-MIR-711

mmu-mir-877

MMU-MIR-877

mmu-mir-669d

MMU-MIR-669D

mmu-mir-713

MMU-MIR-713

mmu-mir-878

MMU-MIR-878

mmu-mir-669e

MMU-MIR-669E

mmu-mir-715

MMU-MIR-715

mmu-mir-879

MMU-MIR-879

mmu-mir-669g

MMU-MIR-669G

mmu-mir-717

MMU-MIR-717

mmu-mir-880

MMU-MIR-880

mmu-mir-669h

MMU-MIR-669H

mmu-mir-719

MMU-MIR-719

mmu-mir-881

MMU-MIR-881

mmu-mir-669j

MMU-MIR-669J

mmu-mir-720

MMU-MIR-720

mmu-mir-883A

MMU-MIR-883A

mmu-mir-669k

MMU-MIR-669K

mmu-mir-721

MMU-MIR-721

mmu-mir-883B

MMU-MIR-883B

mmu-mir-670

MMU-MIR-670

mmu-mir-741

MMU-MIR-741

hsa-mir-885

MIR-885

MIR-671

mmu-mir-742

MMU-MIR-742

hsa-mir-889

MIR-889

mmu-mir-672

MMU-MIR-672

mmu-mir-743a

MMU-MIR-743A

hsa-mir-891a

MIR-891A

mmu-mir-674

MMU-MIR-674

mmu-mir-743b

MMU-MIR-743B

hsa-mir-892b

MIR-892B

mmu-mir-675

MMU-MIR-675

hsa-mir-744

MIR-744

hsa-mir-920

MIR-920

mmu-mir-676

MMU-MIR-676

mmu-mir-744

MMU-MIR-744

hsa-mir-921

MIR-921

mmu-mir-677

MMU-MIR-677

hsa-mir-758

MIR-758

hsa-mir-922

MIR-922

mmu-mir-679

MMU-MIR-679

mmu-mir-758

MMU-MIR-758

hsa-mir-923

MIR-923

mmu-mir-681

MMU-MIR-681

mmu-mir-759

MMU-MIR-759

hsa-mir-924

MIR-924

mmu-mir-682

MMU-MIR-682

mmu-mir-761

MMU-MIR-761

hsa-mir-933

MIR-933

mmu-mir-684-1

MMU-MIR-684-1

mmu-mir-763

MMU-MIR-763

hsa-mir-934

MIR-934

mmu-mir-684-2

MMU-MIR-684-2

mmu-mir-764

MMU-MIR-764

hsa-mir-935

MIR-935

hsa-mir-671

MIR-877

mmu-mir-686

MMU-MIR-686

hsa-mir-766

MIR-766

hsa-mir-936

MIR-936

mmu-mir-688

MMU-MIR-688

hsa-mir-767

MIR-767

hsa-mir-937

MIR-937

mmu-mir-690

MMU-MIR-690

hsa-mir-770

MIR-770

hsa-mir-938

MIR-938

mmu-mir-694

MMU-MIR-694

mmu-mir-770

MMU-MIR-770

hsa-mir-940

MIR-940

mmu-mir-695

MMU-MIR-695

hsa-mir-802

MIR-802

hsa-mir-941

MIR-941

mmu-mir-697

MMU-MIR-697

mmu-mir-802

MMU-MIR-802

hsa-mir-942

MIR-942

mmu-mir-698

MMU-MIR-698

mmu-mir-804

MMU-MIR-804

mmu-mir-1187

MMU-MIR-1187

mmu-mir-699

MMU-MIR-699

mmu-mir-871

MMU-MIR-871

mmu-mir-1188

MMU-MIR-1188

mmu-mir-700

MMU-MIR-700

mmu-mir-872

MMU-MIR-872

mmu-mir-1191

MMU-MIR-1191

mmu-mir-701

MMU-MIR-701

mmu-mir-873

MMU-MIR-873

mmu-mir-1192

MMU-MIR-1192

mmu-mir-702

MMU-MIR-702

hsa-mir-874

MIR-874

mmu-mir-1193

MMU-MIR-1193

mmu-mir-703

MMU-MIR-703

mmu-mir-874

MMU-MIR-874

mmu-mir-1195

MMU-MIR-1195

mmu-mir-704

MMU-MIR-704

mmu-mir-875

MMU-MIR-875

mmu-mir-1197

MMU-MIR-1197

www.cellbiolabs.com

info@cellbiolabs.com

MICRORNA

ANALYSIS

Expression, Control, Reporter Vectors

miRNASelect™ Precursor Clone Collection (Expression Vectors), cont. Product Name

Catalog Number

Product Name

Catalog Number

Product Name

Catalog Number

mmu-mir-1198

MMU-MIR-1198

mmu-mir-1896

MMU-MIR-1896

mmu-mir-1902

MMU-MIR-1902

mmu-mir-1224

MMU-MIR-1224

mmu-mir-1897

MMU-MIR-1897

mmu-mir-1903

MMU-MIR-1903

mmu-mir-1892

MMU-MIR-1892

mmu-mir-1898

MMU-MIR-1898

mmu-mir-1904

MMU-MIR-1904

mmu-mir-1894

MMU-MIR-1894

mmu-mir-1899

MMU-MIR-1899

mmu-mir-1905

MMU-MIR-1905

mmu-mir-1895

MMU-MIR-1895

mmu-mir-1900

MMU-MIR-1900

mmu-mir-1907

MMU-MIR-1907

miRNASelect™ Expression and Control Vectors Our miRNASelect™ Mammalian Expression Vectors provide an easy, efficient method to clone a miRNA precursor from any species. The desired miRNA sequence is cloned into a human ß-globin intron contained within the vector. Two vector formats are available:  

The pEP vector contains a puromycin selection marker The pEGP vector contains a GFP-puromycin fusion to allow selection by either marker

Each expression vector is provided with a null (empty) control vector at no extra charge. Recent Product Citations 1. Esposito, F. et al. (2012). Down-regulation of the miR-25 and miR-30d contributes to the development of anaplastic thyroid carcinoma targeting the polycomb protein EZH2. J. Clin. Endocrinol. Metab. 97:E710-E718. (MIR-EXP-GP-C) 2. Yuan, L. et al. (2012). MicroRNA-212 displays tumor-promoting properties in on-small cell lung cancer cells and targets the hedgehog pathway receptor PTCH1. Mol. Biol. Cell 23:1423-1434. (MIR-EXP-GP-C) Product Name

Size

Catalog Number

miRNASelect™ pEGP-mir Cloning and Expression Vector

100 µL

MIR-EXP-GP-C

miRNASelect™ pEP-mir Cloning and Expression Vector

100 µL

MIR-EXP-C

miRNASelect™ Null Control Vectors Our Null Control vectors have no cloned miRNA sequence. They are useful in conjunction with vectors from our miRNASelect™ Precursor Clone Collection. Recent Product Citations 1. Dar, A. et al. (2013). The role of miR-18b in MDM2-p53 pathway signaling and melanoma progression. J. Natl. Cancer Inst. 105:433-442. (MIR-NULL) 2. Larsen, M. et al. (2014). miRNA-130a regulates C/EBP-epsilon expression during granulopoiesis. Blood 123:1079-1089. (MIR-NULL-GP) 3. Yuan, L. et al. (2012). MicroRNA-212 displays tumor-promoting properties in on-small cell lung cancer cells and targets the hedgehog pathway receptor PTCH1. Mol. Biol. Cell 23:1423-1434. (MIR-NULL-GP)

Product Name

Size

Catalog Number

miRNASelect™ pEGP-mir Null Control Vector

100 µL

MIR-NULL-GP

miRNASelect™ pEP-mir Null Control Vector

100 µL

MIR-NULL

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

miRNA Viral Expression

MICRORNA

ANALYSIS

miRNA Retroviral Expression Vector Our pMXs-miR-GFP/Puro Retroviral Vector allows you to clone a miRNA sequence of interest for packaging into a recombinant retrovirus for delivery into a target cell. Recent Product Citation Mansour, M. et al. (2013). The TAL1 complex targets the FBXW7 tumor suppressor by activating miR-223 in human T cell acute lymphoblastic leukemia. J. Exp. Med. 210:1545-1557. (RTV-017)

For efficient packaging of your miRNA into an MMLV-based retrovirus, use one of our Platinum Retroviral Packaging Cell Lines found on page 64. Product Name

Size

Catalog Number

pMXs-miR-GFP/Puro Retroviral Vector

10 µg

RTV-017

RAPAd® miRNA Adenoviral Expression System RAPAd® Adenoviral Expression Systems produce recombinant adenovirus with substantial reduction in wild-type adenovirus, while doing so in a much shorter 2 week time frame. The RAPAd® miRNA Adenoviral Expression System is specifically designed to deliver miRNA sequences into your target cell.  Virtually No Wild-Type Virus: Backbone vector engineered to produce <300 wild-type plaques per 109 particles, compared with 104-106 WT plaques per 109 VP with most other methods  Faster Virus Production: Virus generated in 2 weeks compared to a few months with traditional methods

Adenovirus Production using the RAPAd® Adenoviral Expression System.

Recent Product Citation Li, P. et al. (2013). MicroRNA-638 is highly expressed in human vascular smooth muscle cells and inhibits PDGF-BB-induced cell proliferation and migration through targeting orphan nuclear receptor NOR1. Cardiovasc. Res. 10.1093/cvr/cvt082. (VPK-253)

For more information on our complete selection of RAPAd® Adenovirus Expression Systems for gene expression studies, please see page 49.

Product Name

Size

Catalog Number

RAPAd® miRNA Adenoviral Expression System

1 Kit

VPK-253

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MICRORNA

ANALYSIS

Reporter System, Knockdown Enhancer

miRNASelect™ Functional Analysis Reporter System The miRNASelect™ Functional Analysis Reporter System provides a simple method for the evaluation of potential targets of miRNA. Binding of miRNA sequences to their suspected targets results in repression of translation. In this system the miRNA target sequence, such as a 3’ UTR, is cloned into the provided pMIRGFP vector in the multiple cloning sites immediately downstream of the GFP gene. If the miRNA is present and binds to the target sequence, translation is repressed and no green fluorescence appears. If the miRNA does not bind the target sequence, GFP translation occurs normally and green fluorescence may be seen.

Assay Principle. If miRNA is present and binds to the 3’ UTR, translation of the GFP gene is repressed, resulting in loss of fluorescence. Product Name

Size

Catalog Number

miRNASelect™ pMIR-GFP Reporter System

1 Kit

MIR-GFP

RNAiBoost™ Reagent Kit for miRNA and siRNA Enhancement RNA interference can occur in the presence of either siRNA or the mature form of miRNA. The RNABoost™ Reagent Kit increases the level of interference in the presence of siRNA. It also increases the rate of processing of pre-miRNA into mature miRNA. Product Name RNAiBoost™ Reagent Kit

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

20 reactions

RNAI-200

100 reactions

RNAI-201

Fax 1-858-271-6514

OXIDATIVE STRESS / DAMAGE

Oxidative Stress Overview

Measuring Oxidative Stress Oxidative stress may be measured using one of three primary methods:   

Measure the reactive oxygen species (ROS) directly Measure the presence of antioxidants Measure the resulting damage to proteins, lipids, DNA or RNA (most reliable)

Use the following table to determine the best oxidative stress assays for your samples. Marker or Type of Damage

Protein Damage (p. 85-89)

Lipid Peroxidation (p. 90-93)

Sample Type Cells Tissues Blood

Protein carbonyl content (PCC)

3-Nitrotyrosine

BPDE Protein Adduct

Advanced Glycation End Products (AGE)

Carboxyethyl Lynsine (CEL)

Carboxymethyl Lynsine (CML)

Methylglyoxal

Advanced Oxidation Protein Products (AOPP)

4-Hydroxynonenal (4-HNE)

Malondialdehyde (MDA)

8-iso-Prostaglandin F2 (8-Isoprostane)

Oxidized Low Density Lipoprotein (OxLDL)

DNA / RNA Damage and Repair (p. 94-100)

8-hydroxydeoxyguanosine (8-OHdG)

Abasic (AP) sites

Aldehyde DNA Damage (Etheno adducts)

BPDE DNA Adduct

Comet Assay

Double-strand DNA breaks

UV DNA Damage (CPD and 6-4PP)

Universal ROS

Hydrogen Peroxide

Cerebrospinal Fluid

Nitric Oxide

Superoxide Dismutase

Catalase

Glutathione

Total Antioxidant Capacity (TAC)

Oxygen Radical Antioxidant Capacity (ORAC)

Food

Hydroxyl Radical Antioxidant Capacity (HORAC)

Food

Cellular Antioxidant Capacity (CAA)

Other

8-hydroxyguanosine (8-OHG)

Reactive Oxygen Species (p. 102-105)

Antioxidants & Antioxidant Capacity (p. 106-110)

Urine

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Saliva

Food

Antioxidant compounds

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Protein Damage

OXIDATIVE STRESS / DAMAGE

Assays and Reagents for Protein Damage Cellular proteins are subject to damage in the presence of reactive oxygen species (ROS). The resulting protein damage may take the form of nitration or oxidation of various amino acid residues, or may result in formation of advanced glycation end products (AGE) or advanced oxidation protein products (AOPP). We have developed unique assays to detect protein damage with higher sensitivity and more user-friendly protocols.

OxiSelect™ Nitrotyrosine Assay Kits and Antibodies Our OxiSelect™ Nitrotyrosine Assay Kits provide a simple method to measure the formation of 3nitrotyrosine in proteins. This assay is available in two formats: a 96-well competitive ELISA and an immunoblot kit. The ELISA format can detect the presence of 3-nitrotyrosine as low as 10 nM. Both kits can detect nitrotyrosine in protein from any species.

Recent Product Citations 1. Toth, P. et al. (2014). Resveratrol treatment rescues neurovascular coupling in aged mice: role of improved cerebromicrovascular endothelial function and downregulation of NADPH oxidase. Am. J. Physiol. Heart Circ. Physiol. 306:H299-H308. (STA305) 2. Li, F.C. et al. (2013). Transition from oxidative stress to nitrosative stress in rostral ventrolateral medulla underlies fatal intoxication induced by organophosphate mevinphos. Toxicol. Sci. 135:202-217. (STA-305) 3. Medina, J.P. et al. (2013). Angiotensin receptor-mediated oxidative stress is associated with impaired cardiac redox signaling and mitochondrial function in insulin-resistant rats. Am. J. Physiol. Heart Circ. Physiol. 305:H599-H607. (STA-305) 4. Kong, X. et al. (2013). Pioglitazone enhances the blood pressure -lowering effect of losartan via synergistic attenuation of angiotensin II-induced vasoconstriction. J. Renin Angiotensin Aldosterone Syst. 10.1177/1170320313489061. (STA-305) 5. Lupachyk, S. et al. (2013). Endoplasmic reticulum stress plays a key role in the pathogenesis of diabetic peripheral neuropathy. Diabetes 62:944-952. (STA-305) 6. Cho, W.K. et al. (2013). IL-13 receptor 2-arginase 2 pathway mediates IL-13-induced pulmonary hypertension. Am. J. Physiol. Lung Cell Mol. Physiol. 304:L112-L124. (STA-305) 7. Montez, P. et al. (2012). Angiotensin receptor blockade recovers hepatic UCP2 expression and aconitase and SDH activities and ameliorates hepatic oxidative damage in insulin resistant rats. Endocrinology 153:5845-5856. (STA-305) 8. Tahrani, A.A. et al. (2012). Obstructive sleep apnea and diabetic neuropathy: a novel association in patients with type 2 diabetes. Am. J. Respir. Crit. Care Med. 186:434-441. (STA-305) 9. Suvakov, S. et al. (2012). Glutathione-S-transferase A1, M1, P1 and T1 null or low-activity genotypes are associated with enhanced oxidative damage among haemodialysis patients. Nephrol. Dial. Transplant. 10.1093/ndt/gfs369. (STA-305) 10.Shevalye, H. et al. (2012). Metanx alleviates multiple manifestations of peripheral neuropathy and increases intraepidermal nerve fiber density in Zucker diabetic fatty rats. Diabetes 61:2126-2133. (STA-305)

Formation of 3-Nitrotyrosine During Oxidative Stress. Product Name

Size

Catalog Number

96 Assays

STA-305

5 x 96 Assays

STA-305-5

Immunoblot/ECL

10 Blots

STA-303

Goat Anti-Nitrotyrosine Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-003

Rabbit Anti-Nitrotyrosine Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-004

Immunoblot/ECL

10 µg

STA-304

Nitrotyrosine ELISA Kit Nitrotyrosine Immunoblot Kit

Protein Tyrosine Nitration Control (Nitrotyrosine-BSA)

Detection Colorimetric

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OXIDATIVE STRESS / DAMAGE

Protein Damage

OxiSelect™ Protein Carbonyl Assay Kits The most common products of protein oxidation in biological samples are the carbonyl derivatives of Pro, Arg, Lys and Thr residues. Such derivatives are chemically stable and serve as markers for oxidative stress in most types of reactive oxygen species. Our OxiSelect™ Protein Carbonyl Assay Kits provide rapid, efficient methods for detection of protein carbonyls. Four assay formats are available: immunoblot, ELISA, fluorometric and spectrophotometric. All formats are suitable for use with purified protein, plasma, serum, or cell lysate samples from any species. Protein Carbonyl ELISA Kit  Sensitive: Detects samples as low as 10 µg/ml  Greater Sample Retention: No concentration or TCA precipitation steps that contribute to sample loss Protein Carbonyl Immunoblot Kit  No Molecular Weight Shift: DNPH derivatization after immunoblotting allows direct comparison of oxidized and non-oxidized protein fingerprints

Assay Principle for the OxiSelect™ Protein Oxidation Immunoblot Kit (STA-308).

Product Name

Detection

Size

Catalog Number

96 Assays

STA-310

5 x 96 Assays

STA-310-5

OxiSelect™ Protein Carbonyl ELISA Kit

Colorimetric

OxiSelect™ Protein Carbonyl Fluorometric Assay

Fluorometric

100 Assays

STA-307

Spectrophotometric

40 Assays

STA-315

OxiSelect™ Protein Carbonyl Immunoblot Kit

Immunoblot/ECL

10 Blots

STA-308

Oxidized Protein Immunoblot Control (Carbonyl-BSA)

Immunoblot/ECL

10 µg

STA-309

OxiSelect™ Protein Carbonyl Spectrophotometric Assay

Recent Product Citations 1. Cui, Z. et al. (2014). Identification of the immunoproteasome as a novel regulator of skeletal muscle differentiation. Mol. Cell Biol. 34:96-109. (STA-308) 2. Pons, D. et al. (2012). Initial activation status of the antioxidant response determines sensitivity to carboplatin/paclitaxel treatment of ovarian cancer. Anticancer Res. 32:4723-4728. (STA-308) 3. Cristovao, A.C. et al. (2012). NADPH oxidase 1 mediates synucleinopathy in Parkinsons’ Disease. J. Neurosci. 32:1446514477. (STA-308) 4. Bitar, M. et al. (2012). Decline in DJ-1 and decreased nuclear translocation of Nrf2 in Fuchs endothelial corneal dystrophy. Invest. Ophthalmol. Vis. Sci. 53:5806-5813. (STA-308) 5. Cannizzo, E. et al. (2012). Age-related oxidative stress compromises endosomal proteostasis. Cell Report 2(1):136-149. (STA308) 6. Satapati, S. et al. (2012). Elevated TCA cycle function in the pathology of diet-induced hepatic insulin resistance and fatty liver. J. Lipid Res. 53:1080-1092. (STA-308) 7. Vulusevic, B. et al. (2014). Glyoxalase-1 overexpression in bone marrow cells reverses defective neovascularization in STZinduced diabetic mice. Cardiovasc. Res. 101:306-316. (STA-310) 8. Dai, D.F. et al. (2013). Global proteomics and pathway analysis of pressure-overload-induced heart failure and its attenuation by mitochondrial-targeted peptides. Circ. Heart Fail. 6:1067-1076. (STA-310) 9. Ungvari, Z. et al. (2013). Testing predictions of the oxidative stress hypothesis of aging using a novel invertebrate model of longevity: the giant clam (Tridacna derasa). J. Gerontol. A Biol. Sci. Med. Sci. 68:359-367. (STA-310) 10.Young, K. et al. (2013). Each to their own: skeletal muscles of different function use different biochemical strategies during aestivation at high temperature. J. Exp. Biol. 216:1012-1024. (STA310) 11.Fishman , J. et al. (2012). Oxidative modification of the intestinal mucus layer is a critical but unrecognized component of trauma hemorrhagic shock-induced gut barrier failure. Am. J. Physiol. Gastrointest. Liver Physiol. 304:G57-G63. (STA-310) 12.Murakami, Y. et al. (2012). Receptor interacting protein kinase mediates necrotic cone but not rod cell death in a mouse model of inherited degeneration. PNAS 10.1073/pnas.1206937109. (STA310) 13.Kang, K.A. et al. (2012). Baicalein inhibits oxidative stressinduced cellular damage via antioxidant effects. Toxic. And Ind. Health 28:412-421. (STA-310) 14.Gannon, A.M. et al. (2012). Cigarette smoke exposure leads to follicle loss via an alternative ovarian cell death pathway in a mouse model. Toxicol. Sci. 125:274-284. (STA-310) 15.Tang, H. et al. (2011). Intrinsic apoptosis in mechanically ventilated human diaphragm: linkage to a novel Fox/FoxO1/Stat3-Bim axis. FASEB J. 25:2921-2936. (STA-310) 16.Robinson, C.K. et al. (2011). A major role for nonenzymatic antioxidant processes in the radioresistance of Halobacterium salinarum. J. Bacteriol. 193:1653-1662. (STA-310)

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Protein Damage

OXIDATIVE STRESS / DAMAGE

OxiSelect™ AOPP Assay Kit Advanced oxidation protein products are toxins created during oxidative stress in patients with diabetes mellitus, atherosclerosis, renal complications, and HIV. Our OxiSelect™ AOPP Assay Kit provides a quick, easy method for assessing AOPP levels.

 Fast: Obtain results in <30 minutes  Sensitive: Detect concentrations as low as 5 µM 1.8 1.6

Recent Product Citations 1. Bloomer, R. et al. (2013). Safety profile of caffeine and 1,3dimethylamylamine supplementation in healthy men. Human and Exp. Toxicol. 10.1177/0960327113475680. (STA-318) 2. Park, S.H. et al. (2012). Effects of neutral pH and low-glucose degradation product-containing peritoneal dialysis fluid on systemic markers of inflammation and endothelial dysfunction: a randomized controlled 1-year follow-up study. Nephrol. Dial. Transp. 27:1191-1199. (STA-318) 3. Anderson, D. et al. (2010). Albumin-based microbubbles bind up -regulated scavenger receptors following vascular injury. J. Biol. Chem. 285:40645-40653. (STA-318) Product Name OxiSelect™ AOPP Assay Kit AOPP-Human Serum Albumin

OD 340nm

1.4 1.2 1 0.8 0.6 0.4 0.2 0

Free HSA

AOPP-HSA

Untreated Human Serum Albumin and AOPP-HSA Positive Control Tested with the OxiSelect™ AOPP Assay Kit. Detection

Size

Catalog Number

Colorimetric

200 Assays

STA-318

N/A

50 µL

STA-319

OxiSelect™ BPDE Protein Adduct ELISA Kit Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, cigarette smoke, and automotive exhaust. They may also be found in some cooked foods. One PAH, benzo(a)pyrene, was the first chemical carcinogen to be discovered. Through a series of enzymatic reactions, benzo(a)pyrene is converted to benzo(a) pyrene 7,8 diol-9,10 epoxide (BPDE) which attacks both proteins and DNA. Our OxiSelect™ BPDE Protein Adduct ELISA Kit provides a convenient method to measure the modification of proteins by BPDE. BPDE-BSA Standard Curve Generated Using the OxiSelect™ BPDE Protein Adduct ELISA Kit.

 Sensitive: Detect concentrations as low as 60 ng/mL  Convenient: Quantify on a standard microplate reader  Versatile: Suitable for use with cell lysates, tissue homogenates, plasma or serum Product Name OxiSelect™ BPDE Protein Adduct ELISA Kit

For information on our BPDE DNA Adduct ELISA Kit, please see page 99.

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-301

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OXIDATIVE STRESS / DAMAGE

Protein Damage

OxiSelect™ Advanced Glycation End Product Kits & Antibodies Advanced glycation end products (AGE) are formed during the Maillard reaction where reducing carbohydrates react with lysine side chains and N-terminal amino groups of various macromolecules, particularly proteins. These AGE products can adversely affect the function of the affected proteins and play a role in atherosclerosis, diabetes, aging and renal disease. Our OxiSelect™ Advanced Glycation End Product Kits are designed for the rapid detection of AGE protein adducts. We offer assays to study generic AGE formation or specific AGE strucutres including N(Carboxyethyl) lysine (CEL), N-(Carboxymethyl) lysine (CML), and methylglyoxal (MG). All kits will detect AGE structures from protein of any species. Advanced Glycation End Products (AGE) Pathways.

OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit Our OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit detects a variety of AGE structures including CML and pentosidine. It does not detect CEL or methylglyoxal (MG). Samples are added to a plate coated with an AGEprotein conjugate. AGE-protein adducts in the sample compete with the AGE-coated plate for antibody binding. High AGE adduct content in a sample results in less binding of the antibody to the plate, producing a low signal.  Sensitive: Detect levels as low as 1 µg/mL of AGEprotein adduct  Versatile: Compatible with cell lysates, plasma, serum, or purified proteins OxiSelect™ AGE Competitive ELISA Kit Standard Curve. Product Name

Detection

OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit Glycoaldehyde-AGE-BSA

Colorimetric N/A

Size

Catalog Number

96 Assays

STA-817

5 x 96 Assays

STA-817-5

100 µg

STA-348

OxiSelect™ N-(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit The OxiSelect™ N-(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit detects CEL protein adducts in a variety of samples including cell lysates, blood samples, and other protein sources. Product Name 

OxiSelect™ N -(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit CEL-BSA

Phone 1-858-271-6500

 Sensitive: Detect levels as low as 100 ng/mL of CEL-protein adduct  Versatile: Compatible with cell lysates, plasma, serum, or purified proteins Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-813

N/A

100 µg

STA-302

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Protein Damage

OXIDATIVE STRESS / DAMAGE

OxiSelect™ N-(Carboxymethyl) Lysine (CML) Assays and Antibodies The OxiSelect™ N-(Carboxymethyl) Lysine (CML) Competitive ELISA Kit detects CML protein adducts in a variety of samples including cell lysates, blood samples, and other protein sources.

 Sensitive: Detect levels as low as 3 ng/mL of CMLprotein adduct with the ELISA kit  Versatile: Compatible with cell lysates, plasma, serum, or purified proteins

CML Protein Adduct Detected in Human Plasma with the OxiSelect™ CML Competitive ELISA Kit. Product Name

Detection

Size

Catalog Number

96 Assays

STA-816

5 x 96 Assays

STA-816-5

OxiSelect™ N-(Carboxymethyl) Lysine (CML) Competitive ELISA Kit

Colorimetric

OxiSelect™ N-(Carboxymethyl) Lysine (CML) Immunoblot Kit

Immunoblot

10 Blots

STA-313

Goat Anti-N-CML Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-013

Rabbit Anti-N-CML Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-014

N/A

100 µg

STA-314

CML-BSA Control

OxiSelect™ Methylglyoxal (MG) Assays and Antibodies  Sensitive: Detect levels as low as 200 ng/mL of MG-protein adduct  Versatile: Compatible with cell lysates, plasma, serum, or purified proteins

The OxiSelect™ Methylglyoxal (MG) Competitive ELISA Kit detects MG protein adducts in a variety of samples including cell lysates, blood samples, and other protein sources. Product Name

Detection

Size

Catalog Number

96 Assays

STA-811

5 x 96 Assays

STA-811-5

Immunoblot/ Immunohistochemistry

100 µg

STA-011

N/A

100 µg

STA-306

Size

Catalog Number

Copper (Cu++) Oxidized Human Low Density Lipoprotein (LDL)

100 µg

STA-214

Malondialdehyde (MDA) Modified Human Albumin

100 µg

STA-210

Malondialdehyde (MDA) Modified Human Apolipoprotein B-100

100 µg

STA-211

Malondialdehyde (MDA) Modified Human Low Density Lipoprotein (LDL)

100 µg

STA-212

Nitrated Human Low Density Lipoprotein (LDL)

100 µg

STA-213

OxiSelect™ Methylglyoxal (MG) Competitive ELISA Kit

Colorimetric

Mouse Anti-Methylglyoxal Monoclonal Antibody MG-BSA

Oxidized / Nitrated Proteins All proteins are provided at a concentration of 1.0 mg/mL. Product Name

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OXIDATIVE STRESS / DAMAGE

Lipid Peroxidation

Assays and Reagents for Lipid Peroxidation Lipid peroxidation is a well-defined mechanism of cellular damage in both animals and plants that occurs during aging and in some disease states. Our OxiSelect™ Lipid Peroxidation Assays allow you to quickly and easily quantify the most common markers and byproducts of lipid peroxidation.

OxiSelect™ MDA (Malondialdehyde) Assays and Antibodies As a common by-product of lipid peroxidation, malondialdehyde (MDA) is a well-accepted marker of oxidative stress. Modification of proteins by MDA can cause structural and functional changes in oxidized proteins. We offer assays and antibodies to measure MDA in a variety of formats. Kits are available to measure total MDA as well as MDA protein adducts specifically.

Note: MDA is most reliably detected in fresh samples, or in samples that have been frozen for a maximum of 1-2 months. For samples stored for longer periods, consider testing other markers of lipid peroxidation such as 4-HNE or 8-isoprostane.

OxiSelect™ TBARS Assay Kit The TBARS assay is a well-established method for screening and monitoring lipid peroxidation via the by-product malondialdehyde (MDA). MDA forms a 1:2 adduct with thiobarbituric acid. Our OxiSelect™ TBARS Assay Kit provides a more user-friendly protocol for quantitation of the MDATBA adduct compared to other commercial assays. This assay detects total MDA, both free and in protein adducts, in a variety of samples including cell and tissue lysates, plasma, and urine. Colorimetric Assay 0.45 0.4

OD 540nm

0.35 0.3 0.25 0.2 0.15 0.1 0.05 0 0

MDA (µM) MDA-TBA Standard Curve Using a Standard Plate Reader. Product Name OxiSelect™ TBARS Assay Kit (MDA Quantitation)

Phone 1-858-271-6500

 Fast: Obtain results in 30 minutes  Sensitive: Smaller reaction volumes require less sample; detect as little as 2 µM  Convenient: 96-well format; no glass tubes are required Recent Product Citations 1. Chang, Q. et al. (2014). Cytochrome P450 2C epoxygenases mediate photochemical stress-induced death of photoreceptors. J. Biol. Chem. 289:8337-8352. 2. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adaptation. J. Gerontol. A Biol. Sci. Med. Sci. 10.1093/gerona/glt122. 3. Fu, L. et al. (2012). Ethyl pyruvate reduces ventilation-induced neutrophil infiltration and oxidative stress. J. Lipid Res. 53:10801092. 4. Brindeiro, C.M. et al. (2012). Tempol prevents altered K+ channel regulation of afferent arteriolar tone in diabetic rat kidney. Hypertension 59:657-664. 5. Joshi, S.G. et al. (2011). Nonthermal dielectric-barrier discharge plasma-induced inactivation involves oxidative DNA damage and membrane lipid peroxidation in Escherichia coli. Antimicrob. Agents Chemother. 55:1053-1062. 6. Kasaikina, M.V. et al. (2011). Roles of the 15-kDa Selenoprotein (Sep15) in redox homeostasis and cataract development revealed by the analysis of Sep15 knockout mice. J. Biol. Chem. 286:33203-33212. 7. Fedeles, B.I. et al. (2011). Chemical genetics analysis of an aniline mustard anticancer reveals complex I of the electron transport chain as a target. J. Biol. Chem. 286:33910-33920. 8. Wojciechowski, P. et al. (2010). Resveratrol arrests and regresses the development of pressure overload- but not volume overload-induced cardiac hypertrophy in rats. J. Nutr. 140:962968. Detection

Size

Catalog Number

Colorimetric or Fluorometric

200 Assays

STA-330

5 x 200 Assays

STA-330-5

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Lipid Peroxidation

OXIDATIVE STRESS / DAMAGE

OxiSelect™ MDA Adduct Assay Kits Our MDA Adduct Assay Kits provide simple methods to measuring these protein adducts in a variety of sample types. The MDA Adduct Competitive ELISA Kit is a sensitive method for the quantitation of MDA in proteins from cells, tissues, or blood. Samples are added to a malondialdehyde protein conjugate-coated plate. The MDA in the sample competes with the MDA on the plate for binding to the primary anti-MDA antibody. A high concentration of MDA in the sample results in little to no antibody binding to the plate, producing a low signal. Our MDA Immunoblot is a convenient method for qualitative measurement of MDA protein adducts.  Sensitive: ELISA kit detects MDA protein adducts as low as 6 pmol/mL  Versatile: Suitable for use with serum, plasma, cell lysates or tissue homogenates Recent Product Citations 1. Montez, P. et al. (2012). Angiotensin receptor blockade recovers hepatic UCP2 expression and aconitase and SDH activities and ameliorates hepatic oxidative damage in insulin resistant rats. Endocrinology 153:5845-5856. (STA-331) 2. Lazrak, A. et al. (2011). Regulation of alveolar epithelial Na+ channels by ERK1/2 in chlorine breathing mice. Am. J. Respir. Cell Mol. Biol. 10.1165/rcmb.2011-0309OC. (STA-331) 3. Zarogiannis, S.G. et al. (2011). Ascorbate and deferoxamine administration post chlorine exposure decrease mortality and lung injury in mice. Am. J. Respir. Cell Mol. Biol. 45:386-392. (STA-331) 4. Hall, J.A. et al. (2010). Absence of thyroid hormone activation during development underlies a permanent defect in adaptive thermogenesis. Endrocrinology 151:4573-4582. (STA-331) 5. Barabutis, N. et al. (2008). Antioxidant activity of growth hormone-releasing hormone antagonists in LNCaP human prostate cancer cell line. PNAS 105:20470-20475. (STA-331) Product Name

Immunoblot of MDA-BSA Control Using the OxiSelect™ MDA Immunoblot Kit. Immunoblot control was electroblotted onto a nitrocellulose membrane, followed by detection with the provided anti-MDA antibody. Detection

Size

Catalog Number

OxiSelect™ MDA Immunoblot Kit

Immunoblot

10 Blots

STA-331

OxiSelect™ MDA Adduct Competitive ELISA Kit

Colorimetric

96 Assays

STA-832

5 x 96 Assays

STA-832-5

N/A

100 µg

STA-333

Detection

Size

Catalog Number

Goat Anti-Malondialdehyde (MDA) Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-031

Rabbit Anti-Malondialdehyde (MDA) Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-032

MDA-BSA

OxiSelect™ MDA Polyclonal Antibodies Product Name

www.cellbiolabs.com

info@cellbiolabs.com

OXIDATIVE STRESS / DAMAGE

Lipid Peroxidation

OxiSelect™ HNE ELISA Kits and Antibodies 4-hydroxynonenal (4-HNE) is a well-known byproduct of lipid peroxidation and is widely accepted as a stable marker for oxidative stress. HNE protein adducts are typically stable when frozen for up to 6 months or more. Our OxiSelect™ HNE Adduct Competitive ELISA Kit provides a simple, user-friendly way to assess HNE adduct formation on lysine, histidine and/or cysteine. Our polyclonal antibodies against HNE recognize HNE adducts to lysine, histidine, and/or cysteine and are suitable for use with Western Blot or ELISA applications.  Sensitive: ELISA kit detects protein adducts as low as 2 µg/mL  Versatile: Suitable for use with serum, plasma, cell lysates or tissue homogenates Standard Curve Generated with the OxiSelect™ HNE Adduct Competitive ELISA Kit.

Product Name

Detection Colorimetric

OxiSelect™ HNE Adduct Competitive ELISA Kit

Size

Catalog Number

96 Assays

STA-838

5 x 96 Assays

STA-838-5

Goat Anti-4-Hydroxynonenal (HNE) Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-034

Rabbit Anti-4-Hydroxynonenal (HNE) Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-035

N/A

100 µg

STA-335

HNE-BSA

OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit (8-isoprostane) 8-iso-Prostaglandin F2 is produced in membrane phospholipids and has been implicated in atherogenesis, rheumatoid arthritis and carcinogenesis. The OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit provides rapid, sensitive detection of 8-iso-PGF2as low as 50 pg/mL. The assay is suitable for quantitation of 8-isoprostane in a variety of sample types including cell and tissue lysates, plasma, serum, and urine.

Product Name OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit

Phone 1-858-271-6500

Recent Product Citations 1. Dugas, T.R. et al. (2014). Hydrogen sulfide cytoprotective signaling is endothelial nitric oxide synthase-nitric oxide dependent. PNAS 111:3182-3187. 2. Karakus, E. et al. (2013). Agomelatine: an antidepressant with new potent hepatoprotective effects on paracetamol-induced liver damage in rats. Human and Experimental Toxicol. 10.0177/0960327112472994. 3. Mollo, R. et al. (2012). Effect of -lipoic acid on platelet reactivity in type 1 diabetic patients. Diabetes Care 35:196-197. 4. Thompson, C.M. et al. (2012). Comparison of the effects of hexavalent chromium in the alimentary canal of F344 rats and B6C3F1 mice following exposure in drinking water: implications for carcinogenic modes of action. Toxicol. Sci. 125:79-90.

Detection Colorimetric

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

96 Assays

STA-337

5 x 96 Assays

STA-337-5

Fax 1-858-271-6514

Lipid Peroxidation

OXIDATIVE STRESS / DAMAGE

OxiSelect™ Human Oxidized LDL ELISA Kits LDL contains a hydrophobic core of various lipids surrounded by one molecule of Apolipoprotein B-100 (ApoB-100), which promotes solubility of the LDL in blood. LDL, often described as “bad” cholesterol, is even more dangerous when it becomes oxidized. Oxidized LDL (OxLDL) is more reactive with surrounding tissues and can collect within the inner lining of arteries.

 Sensitive: Detect as little as 50 ng/mL of MDALDL, 150 ng/mL of CML-LDL, 150 ng/mL of HNELDL, or 100 ng/mL of OxPL-LDL  Quantitative: Compare unknown samples with provided copper oxidized LDL standard

Our OxiSelect™ Human Oxidized LDL ELISA Kits are designed for the detection and quantitation of modified LDL in human plasma or serum. Kits are available to detect MDA-LDL, CML-LDL, or HNE-LDL in either the protein or lipid component of LDL. Our OxPL-LDL kit specifically detects oxidation in the phospholipid component of LDL.

Quantitation of MDA-LDL in Serum and Plasma Samples. Serum and plasma samples were treated with LDL Precipitation Solution. Precipitated LDL pellets were resuspended in 1.6 mL of PBS before further diluting 1:160 in Assay Diluent according to the Assay Protocol.

OxiSelect™ Human Oxidized LDL ELISA Assay Principle.

MDA is the most commonly found damage marker in oxidized LDL, but it can degrade in frozen samples after 1-2 months. CML and HNE, while less commonly found in OxLDL, may be more reliably detectable in samples that have been frozen for several months. Product Name

Detection

Size

Catalog Number

OxiSelect™ Human Oxidized LDL ELISA Kit (CML-LDL Quantitation)

Colorimetric

96 Assays

STA-388

OxiSelect™ Human Oxidized LDL ELISA Kit (HNE-LDL Quantitation)

Colorimetric

96 Assays

STA-389

OxiSelect™ Human Oxidized LDL ELISA Kit (MDA-LDL Quantitation)

Colorimetric

96 Assays

STA-369

OxiSelect™ Human Oxidized LDL ELISA Kit (OxPL-LDL Quantitation)

Colorimetric

96 Assays

STA-358

www.cellbiolabs.com

info@cellbiolabs.com

OXIDATIVE STRESS / DAMAGE

DNA / RNA Damage & Repair

Assays for DNA & RNA Damage and Repair DNA is arguably the most biologically significant target of oxidative and cellular stress. Continuous DNA damage has been implicated in age-related development of various cancers. More recently, RNA damage has been described in conjunction with various neurological diseases including Alzheimer’s and Parkinson’s diseases. We offer a wide range of assays to measure the most common types of DNA and RNA damage in cells.

OxiSelect™ Oxidative DNA Damage ELISA Kit (8-OHdG Quantitation) Among the various types of oxidative DNA damage, 8-hydroxydeoxyguanosine (8-OHdG) is a ubiquitous marker of oxidative stress. 8-OHdG, one of the byproducts of DNA oxidative damage, is physiologically formed and enhanced by chemical carcinogens. Our OxiSelect™ Oxidative DNA Damage ELISA Kit provides a powerful method for rapid quantitation of 8-OHdG in DNA samples. 8-OHdG is easily detectable directly in serum and urine samples, after it is excised and secreted during the DNA repair process. It also may be detected in DNA extracted from cells or tissues of any species, following full DNA digestion into single bases.  Highly Sensitive: Detect as little as 100 pg/ mL of 8-OHdG  Versatile: Suitable for use with urine, serum, and DNA extracted from cells or tissues

2.5

OD 450 nm

2 1.5 1 0.5

0 1/ 64

1/ 32

1/ 16

1/ 80

1/ 40

1/ 20

eg a

tiv

on tr ol di lu t io n di lu t io n di lu t io n di lu t io n di lu t io n di lu t io n

Recent Product Citations 1. Moore, J. et al. (2013). Protection of corneal epithelial stem cells prevents ultraviolet A damage during corneal collagen crosslinking treatment for keratoconus. Br. J. Ophthalmol. 10.1136/ bjophthalmol-2013-303816. 2. Kalghatgi, S. et al. (2013). Disruption of cell-cell junctions and induction of pathological cytokines in the retinal pigment epithelium of light-exposed mice. Sci. Transl. Med. 5:192ra85. 3. James, M.L. et al. (2013). VARA attenuates hyperoxia-induced impaired alveolar development and lung function in newborn mice. Am. J. Physiol. Lung Cell Mol. Physiol. 304:L803-L812. 4. Ravikumar, P. et al. (2013). Klotho protects lung epithelial cells against oxidative DNA damage. FASEB J. 27:722-723. 5. Singh, B. et al. (2013). MicroRNA-93 regulates NRF2 expression and is associated with breast carcinogenesis. Carcinogenesis 10.1093-carcin/bgt026. 6. Singh, B. et al. (2012). Superoxide dismutase 3 is induced by antioxidants, inhibits oxidative DNA damage and is associated with inhibition of estrogen-induced breast cancer. Carcinogenesis 33:2601.2610. 7. Mercer, J.R. et al. (2012). The methyl xanthine caffeine inhibits DNA damage signaling and reactive species and reduces atherosclerosis in ApoE-/- mice. Arterioscler. Thromb. Vasc. Biol. 32:2461-2467. 8. Schutt, F. et al. (2012). Moderately reduced ATP levels promote oxidative stress and debilitate autophagic and phagocytic capacities in human RPE cells. Invest. Ophthalmol. Vis. Sci. 53:5354-5361. 9. Tanabe, K. et al. (2012). Nicorandil as a novel therapy for advanced diabetic nephrophathy in the eNOS-deficient mouse. Am. J. Physiol. Renal Physiol. 302:F1151-F1160. 10.Tzortzaki, E.G. et al. (2012). Oxidative DNA damage and somatic mutations: a link to the molecular pathogenesis of chronic inflammatory airway diseases. Chest 141:1243-1250. 11.Xu, X. et al. (2012). Reactive oxygen species-triggered trophoblast apoptosis is initiated by endoplasmic reticulum stress via activation of caspase-12, CHOP, and the JNK pathway in Toxoplasma gondii infection in mice. Infect. Immun. 80:21212132. 12.Burnham, E. L. (2012). Protandim does not influence alveolar epithelial permeability or intrapulmonary oxidative stress in human subjects with alcohol use disorders. Am. J. Physiol. Lung Cell Mol. Physiol. 302:L688-L699. 13.Pialoux, V. et al. (2011). Losartan abolishes oxidative stress induced by intermittent hypoxia in humans. J. Physiol. 589:55295537.

8-OHdG Levels in a Human Urine Sample. Product Name

Detection

OxiSelect™ Oxidative DNA Damage ELISA Kit (8-OHdG Quantitation)

Phone 1-858-271-6500

Colorimetric

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

96 Assays

STA-320

5 x 96 Assays

STA-320-5

Fax 1-858-271-6514

DNA / RNA Damage & Repair

OXIDATIVE STRESS / DAMAGE

OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation)

Our OxiSelect™ Oxidative RNA Damage ELISA Kit provides a powerful method for rapid quantitation of 8-OHG in urine, serum or cerebrospinal fluid. It also may be used to detect 8-OHG in RNA extracted from cells or tissues of any species. Recent Product Citations 1. Kannan, S. et al. (2012). Dendrimer-based postnatal therapy for neuroinflammation and cerebral palsy in a rabbit model. Sci. Transl. Med. 4:130ra46. 2. Bazin, J. et al. (2011). Targeted mRNA oxidation regulates sunflower seed dormancy alleviation during dry after-ripening. Plant Cell 23:2196-2208.

 Highly Sensitive: Detect as little as 150 pg/ mL of 8-OHG  Versatile: Suitable for use with urine, serum, cerebrospinal fluid, and DNA extracted from cells or tissues 1.6 1.4 1.2

OD 450 nm

Similarly to 8-hydroxydeoxyguanosine (8-OHdG) forming during DNA oxidation, RNA can become oxidized resulting in 8-hydroxyguanosine (8-OHG). Oxidation of RNA has been implicated in a number of neurological diseases including Alzheimer’s and Parkinson’s diseases.

1 0.8 0.6 0.4 0.2 0 0.01

0.1

100

8-OHG (ng/mL) Standard Curve Generated with the OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG). Product Name

Detection

OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation)

Colorimetric

Size

Catalog Number

96 Assays

STA-325

5 x 96 Assays

STA-325-5

OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine Quantitation) Various reactive nitrogen species (RNS) including peroxynitrite and nitrogen oxides can form during pathophysiological conditions. These RNS can nitrate guanine bases to form 8-nitroguanine in both DNA and RNA. Nitrosative damage to DNA and RNA is a significant contributor to the age-related development of major inflammation-related diseases as well as colon, breast, and prostate cancers. Our OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit provides a simple method for rapid quantitation of 8-nitroguanine in urine, serum or plasma samples. The assay measures total 8-nitroguanine from both DNA and RNA combined. Standard Curve Generated with the OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine). Product Name OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine Quantitation)

Detection Colorimetric

www.cellbiolabs.com

Size

Catalog Number

96 Assays

STA-825

5 x 96 Assays

STA-825-5

info@cellbiolabs.com

OXIDATIVE STRESS / DAMAGE

DNA / RNA Damage & Repair

OxiSelect™ Oxidative DNA Damage Quantitation Kit (AP Sites)

Our OxiSelect™ Oxidative DNA Damage Quantitation Kit provides a simple, user-friendly method for measuring AP sites in DNA. The assay uses an aldehyde reactive probe (ARP) which specifically reacts with an aldehyde group on the open ring of the AP site, followed by labeling with Biotin and subsequent detection by Streptavidin-enzyme conjugate.

1.8 1.6 1.4

OD 450 nm

Oxidative DNA Damage can manifest in the formation of apurinic or apyrimidinic (AP) sites, also known as loss of bases. Spontaneous base loss, if unrepaired, can inhibit transcription and may be mutagenic.

1.2 1 0.8 0.6 0.4 0.2 0 0

AP Sites per 100,000bp

 Highly Sensitive: Detect as few as 4-40 AP sites in 105 bp of DNA  Versatile: Suitable for use with genomic DNA from cells or tissues  Quantitative: Kit includes both oxidized and reduced DNA standards for absolute quantitation Product Name OxiSelect™ Oxidative DNA Damage Quantitation Kit (AP Sites)

Standard Curve Generated with the OxiSelect™ Oxidative DNA Damage Quantitation Kit (STA-324). Recent Product Citations 1. Messaoudi, N. et al. (2013). Global stress response in a prokaryotic model of DJ-1-associated Parkinsonism. J. Bacteriol. 195:1167-1178. 2. Zaika, E. et al. (2011). P73 protein regulates DNA damage repair. FASEB J. 25:4406-4414. Detection

Size

Catalog Number

Colorimetric

50 Assays

STA-324

OxiSelect™ DNA Double-Strand Break Assay Double-strand breaks (DSB) are among the most dangerous types of DNA damage within cells. An early cellular response is phosphorylation of the histone variant H2AX at the site of the DSB. This triggers a cascade of events and appears to play a role in recruitment of repair factors to the damaged sites. Our OxiSelect™ DNA Double-Strand Break Staining Kit provides an easy-to-use method for detecting DNA breaks. The kit utilizes simple immunofluorescence staining of the phosphorylated histone H2AX.

 Fast: See staining results in about 3 hours  Positive Control: DNA Double-strand break inducer included in kit Product Name OxiSelect™ DNA Double-Strand Break Staining Kit

Phone 1-858-271-6500

DNA Double-Strand Break Formation in A549 Cells. A549 cells were seeded at 50,000 cells/well overnight. Immunofluorescence staining was then performed according to the assay protocol. (A) Untreated cells. (B) Cells treated with 100 µM etoposide for one hour.

Detection

Size

Catalog Number

Immunofluorescence

100 Assays

STA-321

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

DNA / RNA Damage & Repair

OXIDATIVE STRESS / DAMAGE

OxiSelect™ Comet Assays (Single Cell Gel Electrophoresis) DNA damage can result from a variety of intracellular and extracellular stimuli, and can manifest in a variety of mutations to the DNA including base modifications, missing bases and single-stranded or double-stranded breaks. Traditionally the comet assay, or single cell gel electrophoresis (SCGE), has been used as a wellpublished, high-level screening tool to measure DNA damage in single cells. Our OxiSelect™ Comet Assay Kits provide a quick, easy method to screen for DNA damage at a macro level. Our OxiSelect™ Comet Assay Slides have been specially treated for adhesion of low-melting agarose used in the assay. Damaged DNA moves farther in electrophoresis than intact DNA, causing a “tail” to form upon visualization under a fluorescence microscope. Recent Product Citations 1. Luo, Y. et al. (2013). SMC1-mediated intra-S-phase arrest facilitates bocavirus DNA replication. J. Virol. 87:4017-4032. (STA-350, STA-351) 2. Li, Y.J. et al. (2012). Gold nanoparticles as a platform for creating a multivalent poly-SUMO chain inhibitor that also augments ionizing radiation. PNAS 109:4092-4097. (STA350, STA-351) 3. Robin, T.P. et al. (2012). EWS/FLI1 regulates EYA3 in Ewing Sarcoma via modulation of miRNA-708, resulting in increased cell survival and chemoresistance. Mol. Cancer Res. 10:10981108. (STA-350, STA-351) 4. Choi, S.K. et al. (2012). Poly(ADP-ribose) polymerase 1 inhibition improves coronary arteriole function in type 2 diabetes mellitus. Hypertension 59:1060-1068. (STA-350, STA-351) 5. Tyagi, A. et al. (2011). Resveratrol selectively induces DNA damage, independent of Smad4 expression, in its efficacy against human head and neck squamous cell carcinoma. Clin. Cancer Res. 17:5402-5411. (STA-355) Product Name

OxiSelect™ 3-Well Comet Assay Kit

OxiSelect™ 3-Well Comet Assay Slides

Assay Principle for the OxiSelect™ Comet Assay Kit.

Etoposide Treatment of Jurkat Cells. Jurkat cells were either untreated (left) or treated with etoposide (right) prior to performing the OxiSelect™ Comet Assay. Detection

Light Microscopy

OxiSelect™ 96-Well Comet Assay Kit

Light Microscopy

OxiSelect™ 96-Well Comet Assay Slides

Light Microscopy

OxiSelect™ Comet Assay Control Cells (includes positive and negative controls)

N/A

www.cellbiolabs.com

Size

Catalog Number

15 Wells

STA-350

75 Wells

STA-351

5 x 75 Wells

STA-351-5

5 Slides

STA-352

25 Slides

STA-353

125 Slides

STA-353-5

96 Wells

STA-355

5 x 96 Wells

STA-355-5

1 Slide

STA-356

5 Slides

STA-356-5

1 Set

STA-354

info@cellbiolabs.com

OXIDATIVE STRESS / DAMAGE

DNA / RNA Damage & Repair

OxiSelect™ UV-Induced DNA Damage Assays

Our OxiSelect™ UV-Induced DNA Damage Assays conveniently measure the formation of either CPD or 6-4PP in intact cells. Kits for each marker are available in three formats: an ELISA for DNA extracted from cells or tissues, a Cell-Based ELISA, and a Cellular Immunostaining kit.

Recent Product Citations 1. Zirkin, S. et al. (2013). The PIM-2 kinase is an essential component of the ultraviolet damage response that acts upstream to E2F-1 and ATM. J. Biol. Chem. 288:21770-21789. (STA322) 2. Burgess, H.M. et al. (2011). Nuclear relocalisation of cytoplasmic poly(A)-binding proteins PABP1 and PABP4 in response to UV irradiation reveals mRNA-dependent export of metazoan PABPs. J. Cell Sci. 124:3344-3355. (STA-322)

1.6 1.4 1.2 OD 450nm

Absorption of ultraviolet radiation can damage DNA by the formation of pyrimidine dimers. The two most common forms of pyrimidine dimers are cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) pyrimidone photoproducts (6-4PP).

1.0 0.8 0.6 0.4 0.2 0.0 Secondary Antibody Alone

UV-Induced DNA Damage in HeLa Cells Treated with Ultraviolet Light for 30 Minutes and Visualized with the OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (CPD).

UV Treated

Untreated

UV-Induced DNA Damage in HeLa Cells as Detected with the OxiSelect™ Cellular UV-Induced DNA Damage ELISA (CPD).

OxiSelect™ UV-Induced DNA Damage ELISA Kits, for extracted DNA Product Name

Detection

Size

Catalog Number

OxiSelect™ UV-Induced DNA Damage ELISA Combo Kit (CPD/6-4PP)

Colorimetric

96 Assays

STA-322-C

OxiSelect™ UV-Induced DNA Damage ELISA Kit (CPD Quantitation)

Colorimetric

96 Assays

STA-322

5 x 96 Assays

STA-322-5

OxiSelect™ UV-Induced DNA Damage ELISA Kit (6-4PP Quantitation)

Colorimetric

96 Assays

STA-323

5 x 96 Assays

STA-323-5

OxiSelect™ Cellular UV-Induced DNA Damage Staining Kits, for intact cells Product Name

Detection

OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kit (CPD)

Colorimetric

OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kit (6-4PP)

Colorimetric

Size

Catalog Number

96 Assays

STA-326

5 x 96 Assays

STA-326-5

96 Assays

STA-328

OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kits, for intact cells Product Name

Detection

Size

Catalog Number

OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (CPD)

Fluorescence Microscopy

96 Assays

STA-327

OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (6-4PP)

Fluorescence Microscopy

96 Assays

STA-329

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

DNA / RNA Damage & Repair

OXIDATIVE STRESS / DAMAGE

OxiSelect™ Aldehyde-Induced DNA Damage Assays (Etheno Adducts) Oxidation of phospholipids can lead to the formation of lipid hydroperoxides. These resulting short-lived hydroperoxides can either be converted to inert fatty acid alcohols, or can react with metals to form aldehydes such as malondialdehyde (MDA), 4-hydroxynonenal (HNE), acrolein, and crotonaldehyde. These aldehydes (which can also be formed through exposure to carcinogenic substances such as urethane or vinyl chloride) can damage DNA resulting in the formation of various etheno adducts, including 1,N6-ethenodeoxyadenosine and 3,N4ethenodeoxycytidine. The presence of these bases can lead to base pair substitution mutations. Our OxiSelect™ Aldehyde-Induced DNA Damage Assays conveniently measure the formation of either 1,N6-ethenodeoxyadenosine (ethenoadenosine) or 3,N4-ethenodeoxycytidine (ethenocytidine) in DNA extracted from cells or tissues. In addition, we offer a convenient combination kit that can measure both etheno bases in separate wells of the same plate. Etheno Base Structures that Form Adducts with DNA During Oxidative Stress. Product Name

Detection

Size

Catalog Number

OxiSelect™ Aldehyde-Induced DNA Damage ELISA Combo Kit (Ethenoadenosine / Ethenocytidine Quantitation)

Colorimetric

96 Assays

STA-820-C

OxiSelect™ Aldehyde-Induced DNA Damage ELISA Kit (Ethenoadenosine Quantitation)

Colorimetric

96 Assays

STA-820

OxiSelect™ Aldehyde-Induced DNA Damage ELISA Kit (Ethenocytidine Quantitation)

Colorimetric

96 Assays

STA-821

OxiSelect™ BPDE DNA Adduct ELISA Kit Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, cigarette smoke, and automotive exhaust. One PAH, benzo(a)pyrene, was the first chemical carcinogen to be discovered. Through a series of enzymatic reactions, benzo(a)pyrene is converted to benzo(a)pyrene 7,8 diol-9,10 epoxide (BPDE) which attacks both proteins and DNA. Our OxiSelect™ BPDE DNA Adduct ELISA Kit provides a convenient method to measure BPDE adducts in DNA extracted from cells or tissues.  Sensitive: Detect concentrations as low as 30 ng/mL  Convenient: Quantify on a standard microplate reader Recent Product Citation Chiu, C.Y. et al. (2014). Low-dose benzo(a)pyrene and its epoxide metabolite inhibit myogenic differentiation in human skeletal muscle-derived progenitor cells. Toxicol. Sci. 10.1093/toxsci/ kfu003. Product Name OxiSelect™ BPDE DNA Adduct ELISA Kit

BPDE-DNA Standard Curve Generated Using the OxiSelect™ BPDE DNA Adduct ELISA Kit.

For information on our BPDE Protein Adduct ELISA Kit, please see page 87. Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-357

www.cellbiolabs.com

info@cellbiolabs.com

OXIDATIVE STRESS / DAMAGE

DNA / RNA Damage & Repair

Checkpoint Kinase Activity Assays Checkpoint kinases, specifically CHK1 and CHK2, are activated in response to DNA damage to subsequently phosphorylate Cdc25C prior to mitosis, which prompts cell cycle arrest. Mutation of these checkpoint kinases can ultimately lead to decreased DNA repair. Our Checkpoint Kinase Activity Assays allow you to conveniently measure the activity of CHK1 and CHK2. The assays use recombinant Cdc25C as a checkpoint kinase substrate. Phosphorylated Cdc25C (Ser216) is detected using a phospho-specific antibody. Checkpoint Kinase Activitiy Assays are available in two formats: a Western blot assay and a 96-well plate-based activity assay. Product Name

Detection

Size

Catalog Number

Checkpoint Kinase Activity Immunoblot Kit

Immunoblot

20 Assays

STA-413

96 Assays

STA-414

96-Well Checkpoint Kinase Activity Assay Kit

Colorimetric

5 x 96 Assays

STA-414-5

Global DNA Methylation ELISA Kit DNA methylation is an epigenetic change shown to be associated with nearly every biological process. In mammalian cells, DNA methylation is found predominantly at CpG dinucleotides; however, in certain cases such as embryonic stem cells it may also be found in non-CpG contexts. Due to the important role of DNA methylation in maintaining genomic stability, deregulation of DNA methylation is associated with various diseases including cancer. Our Global DNA Methylation and Hydroxymethylation Assays provide a convenient, accurate way to quantify 5’-methyl-2’-deoxycytidine (5MedCyd) and 5-hydroxymethylcytosine respectively. Unknown samples are compared with a standard provided with each kit.

 Sensitive: Detect as little as 15 nM of 5MedCyd  Versatile: Suitable for use with any isolated DNA as well as urine samples  Convenient: Quantify on a standard microplate reader

Methylated DNA Standard Curve Generated with the Global DNA Methylation ELISA Kit.

5MedCyd Levels in Human Urine Sample as Measured with the Global DNA Methylation ELISA Kit.

Product Name Global DNA Methylation ELISA Kit (5’-methyl-2’-deoxycytidine Quantitation)

100

Phone 1-858-271-6500

Detection Colorimetric

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

96 Assays

STA-380

5 x 96 Assays

STA-380-5

Fax 1-858-271-6514

Hypoxia Assays

OXIDATIVE STRESS / DAMAGE

HIF-1 Alpha DNA Binding Activity Assay Kit Cell hypoxia, or low oxygen condition, is a normal physiological response to certain body stressors such as high altitudes, but it also can be a symptom of pathological conditions. In some cases hypoxia may contribute to the inducement of oxidative stress. In response to hypoxic conditions, the hypoxia-inducible factor 1 transcriptional activator complex (HIF-1) plays a role in activating several hypoxia-responsive genes such as erythropoietin and VEGF. During hypoxia, the alpha subunit of HIF-1 accumulates and translocates from the cytosol to the nucleus, where it dimerizes with the beta subunit and becomes transcriptionally active. It then binds transcriptional coactivators to induce gene expression. The HIF-1 Alpha DNA Binding Activity Assay Kit is an ELISA-based assay to detect activated HIF-1. Active HIF-1 complex is captured on a double-stranded oligo containing a hypoxic response element (HRE) that is attached to the plate. Detection is then performed with a primary antibody followed by an HRPconjugated secondary antibody. The assay will detect HIF-1 complexes from human, mouse or rat samples. Product Name HIF-1 Alpha DNA Binding Activity Assay Kit

Size

Catalog Number

Colorimetric

96 Assays

CBA-282

Detection of Nuclear HIF-1 Alpha. HeLa cells were incubated in the presence or absence of 0.2 mM DFO for 4 hours at 37ºC. Nuclear extracts were prepared using the Nuclear/Cytosolic Fractionation Kit. HIF-1 Alpha levels were measured in untreated (blue bars) and treated (red bars) extracts according to the Assay Protocol. Detection

Size

Catalog Number

HIF-1 Alpha Sandwich ELISA Kit

Colorimetric

96 Assays

CBA-280

HIF-1 Alpha Cell Based ELISA Kit

Chemiluminescent

96 Assays

CBA-281

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101

OXIDATIVE STRESS / DAMAGE

ROS Assays

Reactive Oxygen Species Assays Reactive oxygen species (ROS) such as superoxide and hydrogen peroxide are continually produced during metabolic processes. Excess ROS can lead to cellular injury in the form of damaged DNA, lipids and proteins. We offer assays for quantitation of various reactive oxygen species, in both in vitro and intracellular formats.

OxiSelect™ Intracellular ROS Assay Kit The OxiSelect™ Intracellular ROS Assay Kit measures the activity of hydroxyl, peroxyl, and other reactive oxygen species. The assay uses the cell-permeable fluorogenic probe DCFH-DA, which diffuses into cells and is deacetylated into the non-fluorescent DCFH. In the presence of ROS, the DCFH is oxidized into highly fluorescent DCF. Fluorescence is quantified on a fluorometric plate reader.  Sensitive: Detect concentrations as little as 10 pM  Fast: Entire protocol takes about one hour Recent Product Citations 1. Koontz, J. et al. (2014). Competition through dimerization between antiapoptotic and proapoptotic HS-1-associated protein X1 (Hax-1). J. Biol. Chem. 289:3468-3477. 2. Kim, E.Y. et al. (2013). NOX2 interacts with podocyte TRPC6 channels and contributes to their activation by diacylglycerol: essential role of podocin in formation of this complex. Am. J. Physiol. Cell Physiol. 305:C960-C971. 3. Abe, Y. et al. (2013). TGF-ß1 stimulates mitochondrial oxidative phosphorylation and generation of reactive oxygen species in cultured mouse podocytes, mediated in part by the mTOR pathway. Am. J. Phyisol. Renal Physiol. 305:F1477-F1490. 4. He, Q. et al. (2013). Tafazzin knockdown interrupts cell cycle progression in cultured neonatal ventricular fibroblasts. Am. J. Physiol. Heart Circ. Physiol. 305:H1332-H1343. 5. Kokkinaki, M. et al. (2013). Klotho regulates retinal pigment epithelial functions and protects against oxidative stress. J. Neurosci. 33: 16346-16359. 6. Hagan, C. et al. (2013). A common docking domain in progesterone receptor-B links DUSP6 and CK2 signaling to proliferative transcriptional programs in breast cancer cells. Nucleic Acids Res. 10.1093/nar/gkt706. 7. Teng, H. et al. (2013). Oxygen-sensitive mitochondrial accumulation of cystathione ß-synthase mediated by lon protease. PNAS 110:12679-12684. 8. Wang, H. et al. (2012). p53-induced gene 3 mediates cell death induced by glutathione peroxidase 3. J. Biol. Chem. 287:1689016902. 9. Druz, A. et al. (2012). Glucose depletion activates MMU-mir466h-5p expression through oxidative stress and inhibition of histone deacetylation. Nucleic Acids Res. 10.1093/nar/gks452. 10.Montalvo-Ortiz, B.L. et al. (2012). Characterization of EHOP016, novel small molecule inhibitor of Rac GTPase. J. Biol. Chem. 287:13228-13238. Assay Principle for the OxiSelect™ Intracellular ROS Assay. Product Name OxiSelect™ Intracellular ROS Assay Kit

102

Phone 1-858-271-6500

Detection Fluorometric

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

96 Assays

STA-342

5 x 96 Assays

STA-342-5

Fax 1-858-271-6514

ROS Assays

OXIDATIVE STRESS / DAMAGE

OxiSelect™ In Vitro ROS/RNS Assay Kit Free radicals and related reactive oxygen species (ROS) and reactive nitrogen species (RNS) can appear in the body both inside and outside the cell. Until recently it has been difficult to detect ROS and RNS outside of intact cells. The OxiSelect™ In Vitro ROS/RNS Assay Kit allows you to measure ROS and RNS formation in various body fluids including urine, serum and plasma. It is also useful for testing cell lysates, tissue homogenates, and cell culture supernatants.

 Sensitive: Detect concentrations as little as 10 pM for DCF or 40 nM for hydrogen peroxide  Fast: Entire protocol takes about one hour  Versatile: Suitable for a wide variety of sample types including urine, serum, plasma, cell lysates, tissue homogenates and cell culture supernatants

The assay universally measures reactive oxygen and reactive nitrogen species that may include hydrogen peroxide, nitric oxide, peroxynitrite, peroxyl radicals, and others. The assay principle is similar to our Intracellular ROS Assay (previous page), except that the chemistry is modified to allow detection of ROS outside the cell. Fluorescence is quantified on a fluorometric plate reader.

Recent Product Citations 1. Liu, X. et al. (2013). Epoxyeicosatrienoic acids prevent cisplatininduced renal apoptosis through a p38 mitogen-activated protein kinase-regulated mitochondrial pathway. Mol. Pharmacol. 84:925-934. 2. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adaptation. J. Gerontol. A Biol. Sci. Med. Sci. 10.1093/gerona/glt122. 3. Xiao, D. et al. (2013). Estrogen normalizes perinatal nicotineinduced hypertensive responses in adult female rat offspring. Hypertension 61:1246-1252. 4. Wang, W. et al. (2012). Mono-(2-ethylhexyl) phthalate induces oxidative stress and inhibits growth of mouse ovarian antral follicles. Biol. Reprod. 87:152. 5. Ju, D. J. et al. (2012). Ethyl pyruvate ameliorates albuminuria and glomerular injury in the animal model of diabetic nephropathy. Am. J. Phyisol. Renal Physiol. 302:F606-F613. 6. Momi, S. et al. (2012). Nitric oxide enhances the antiinflammatory and anti-atherogenic activity of atorvastatin in a mouse model of accelerated atherosclerosis. Cardiovasc. Res. 10.1093/cvr/cvs100. 7. Patterson, A.J. et al. (2011). Hypoxia-derived oxidative stress mediates epigenetic repression of PKC epsilon gene in foetal rat hearts. Cardiovasc. Res. 10.1093/cvr/cvr322. 8. Rathnasamy, G. et al. (2011). Iron and iron regulatory proteins in amoeboid microglial cells are linked to oligodendrocyte death in hypoxic neonatal rat periventricular white matter through production of proinflammatory cytokines and reactive oxygen/nitrogen species. J. Neurosci. 31:17982-17995.

Product Name OxiSelect™ In Vitro ROS/RNS Assay Kit

Assay Principle for the OxiSelect™ In Vitro ROS/RNS Assay.

Detection Fluorometric

www.cellbiolabs.com

Size

Catalog Number

96 Assays

STA-347

5 x 96 Assays

STA-347-5

info@cellbiolabs.com

103

OXIDATIVE STRESS / DAMAGE

ROS Assays

OxiSelect™ Hydrogen Peroxide Assay, Colorimetric Hydrogen peroxide is one of the most prevalent and most stable of the various reactive oxygen species. The half-life of hydrogen peroxide is significantly longer than that of most ROS, making it easier to detect in many sample types. Our OxiSelect™ Hydrogen Peroxide Assay Kit provides a simple method for quantitation of hydrogen peroxide. This colorimetric assay measures the oxidation of ferrous (Fe2+) ions to ferric (Fe3+) ions in the presence of peroxides. The ferric ions form a complex with a provided dye which may be read on a standard microplate reader. The assay may be run with either aqueous phase or lipid phase samples.

 Sensitive: Detect as little as 1 µM  Fast: Easy 30-90 minute incubation, depending on sample type  Versatile: Suitable for plasma, serum, urine, and cell culture supernatants*

*For the testing of hydrogen peroxide in cells and tissues, please see our OxiSelect™ Hydrogen Peroxide / Peroxidase Assay Kit below. Product Name OxiSelect™ Hydrogen Peroxide Assay Kit

Detection

Size

Catalog Number

Colorimetric

500 Assays

STA-343

OxiSelect™ Hydrogen Peroxide / Peroxidase Assay, Fluorometric Our OxiSelect™ Hydrogen Peroxide / Peroxidase Assay Kit provides a convenient plate-based method for quantitation of hydrogen peroxide or peroxidases in a wide variety of sample types. This fluorometric assay uses a fluorogenic probe which is converted from a non-fluorescent to a fluorescent state in the presence of peroxides and is catalyzed by peroxidases.

 Sensitive: Detect as little as 50 nM  Fast: Easy 30 minute incubation  Versatile: Measure either hydrogen peroxide or peroxidase in plasma, serum, urine, cell culture supernatants, cell lysates and tissue homogenates

The kit includes both a hydrogen peroxide standard and a peroxidase standard for quantitative results with either target.

Recent Product Citations 1. Kim, E.Y. et al. (2012). Sustained activation of N-methyl-Daspartate receptors in podocytes leads to oxidative stress, mobilization of transient receptor potential canonical 6 channels, nuclear factor of activated T cells activation, and apoptotic cell death. Mol. Pharmacol. 82:728-737. 2. Kim, E.Y. et al. (2012). Insulin increases surface expression of TRPC6 channels in podocytes: role of NADPH oxidases and reactive oxygen species. Am. J. Phyisol. Renal Physiol. 302:F298-F307. Standard Curve Generated with the OxiSelect™ Hydrogen Peroxide/Peroxidase Assay (Fluorometric). Product Name OxiSelect™ Hydrogen Peroxide/Peroxidase Assay Kit

104

Phone 1-858-271-6500

Detection

Size

Catalog Number

Fluorometric

500 Assays

STA-344

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

ROS Assays

OXIDATIVE STRESS / DAMAGE

OxiSelect™ Intracellular Nitric Oxide Assay Kit Nitric oxide (NO) is a progenitor of various reactive nitrogen species (RNS) in conjunction with superoxide anions via nitric oxide synthase (NOS). It plays a role in vascular diseases, diabetes, atherosclerosis, inflammatory diseases and cancer. Because of its short half-life, nitric oxide is often difficult to detect directly. The OxiSelect™ Intracellular Nitric Oxide Assay Kit allows direct detection of NO in intact cells. A cellpermeable fluorogenic probe is added to cells; upon treatment to induce oxidative stress, nitric oxide that is generated within the cell binds to the probe producing a bright fluorescent signal. Results may be visualized under a fluorescence microscope or quantified in a 96-well fluorescence plate reader.

Product Name

 Direct detection: Probe binds directly to nitric oxide, not to by-products such as nitrate and nitrite  Sensitive: Detect as little as 3 nM  Versatile: Read results as endpoint or time course (kinetic) in a fluorescence plate reader, or visualize under a fluorescence microscope

Induction of NOS in RAW 264.7 Cells. Cells were seeded in a 96-well plate at 100,000 cells/well. Cells were uninduced (left) or induced with 50 ng/mL LPS and 10 ng/mL IFN(right) for 20 hours at 37ºC. Detection

OxiSelect™ Intracellular Nitric Oxide (NO) Assay Kit

Fluorometric

Size

Catalog Number

96 Assays

STA-800

5 x 96 Assays

STA-800-5

OxiSelect™ In Vitro Nitric Oxide Assay Kits Nitric oxide (NO) is difficult to detect directly in vitro due to its short half-life. It is therefore common to measure nitric oxide formation by detection of its final oxidized products, nitrate and nitrite. The OxiSelect™ In Vitro Nitric Oxide Assay Kits provide a convenient plate-based method for the quantitation of nitrate and nitrite in a variety of sample types. First, nitrate is reduced to nitrite. Then total nitrite is measured by the addition of a Griess Reagent (for colorimetric detection) or a fluorometric probe (for fluorescence detection). Results are then quantified in a 96-well plate reader. OxiSelect™ In Vitro Nitric Oxide Assay Kits are suitable for use with serum, plasma, urine, saliva, cell lysates, and culture media. Assay Principle for the OxiSelect™ In Vitro Nitric Oxide (Nitite / Nitrate) Assay, Fluorometric Format. Product Name

Detection Colorimetric

OxiSelect™ In Vitro Nitric Oxide (Nitrite / Nitrate) Assay Kit Fluorometric

www.cellbiolabs.com

Size

Catalog Number

100 Assays

STA-802

5 x 100 Assays

STA-802-5

100 Assays

STA-801

5 x 100 Assays

STA-801-5

info@cellbiolabs.com

105

OXIDATIVE STRESS / DAMAGE

Antioxidant Assays

Antioxidant Assays ROS generation is normally counterbalanced by the action of antioxidant enzymes and other redox molecules. We offer two types of assays for antioxidant quantitation:  

Assays to quantify the presence or activity of antioxidant molecules Assays to determine the antioxidant capacity of biomolecules

OxiSelect™ Catalase Activity Assay Kits

Our OxiSelect™ Catalase Activity Assay Kits provide a quick, user-friendly protocol to monitor catalase activity from a variety of sample types. Kits are available with either colorimetric or fluorometric detection.  Sensitive: Detect as little as 1.25 units/mL (colorimetric) or 50 mU/mL (fluorometric)  Fast: Obtain results in less than 30 minutes  Versatile: Suitable for use with whole blood, plasma, serum, cell lysates or tissue homogenates  High Throughput: 96-well format Recent Product Citation Costantini, D. et al. (2013). Loss of integration is associated with reduced resistance to oxidative stress. J. Exp. Biol. 216:22132220. (STA-341)

1.8 1.6 1.4 1.2

OD 540nm

Catalase is a ubiquitous enzyme that destroys hydrogen peroxides formed during oxidative stress. Since hydrogen peroxides have a longer half-life than most free radicals and can make up a large portion of all reactive oxygen species, the ability to remove hydrogen peroxides can be extremely important at combating oxidative stress.

1 0.8 0.6 0.4 0.2 0 0

100

Catalase (U/mL) Standard Curve Generated with the OxiSelect™ Catalase Activity Assay Kit (Colorimetric).

Assay Principle for the OxiSelect™ Catalase Acitivity Assays. Catalase present in samples converts hydrogen peroxide into water and oxygen (Reaction 1). Any remaining hydrogen peroxide that is not converted reacts with a colorimetric or fluorometric probe in the presence of horseradish peroxidase (Reaction 2) to produce a color or fluorescence which is measured in a plate reader. Product Name OxiSelect™ Catalase Activity Assay Kit

106

Phone 1-858-271-6500

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-341

Fluorometric

96 Assays

STA-339

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Antioxidant Assays

OXIDATIVE STRESS / DAMAGE

OxiSelect™ Superoxide Dismutase Activity Assay Superoxide dismutase (SOD), which catalyzes the dismutation of the superoxide anion into hydrogen peroxide and molecular oxygen, is one of the most important antioxidant enzymes. The OxiSelect™ Superoxide Dismutase Activity Assay uses a xanthine/xanthine oxidase (XOD) system to generate superoxide anions and a chromagen to produce a water-soluble dye upon reduction by the superoxide anions.

 Sensitive: Detect as little as 0.6 units/mL  Fast: Obtain results in about 2 hours  Versatile: Suitable for use with urine, serum, cells or tissue samples

OxiSelect™ Superoxide Dismutase Activity Assay Principle. Superoxide anions generated by a Xanthine/Xanthine Oxidase system are detected with the provided chromagen. SOD reduces superoxide concentrations, so higher SOD concentrations result in a decreased signal.

1 0.9

OD 490nm

0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 0.001

0.01

0.1

100

SOD (Units)

Recent Product Citations 1. Wysocki, J. et al. (2014). ACE2 deficiency increases NADPHmediated oxidative stress in teh kidney. PHY2 2:e00264. 2. Yin, J. et al. (2014). Development of an antioxidant system after early weaning in piglets. J. Anim. Sci. 92:612-619. 3. Gong, E.J. et al. (2013). Low-dose-rate radiation exposure leads to testicular damage with decreases in DNMT1 and HDAC1 in the murine testis. J. Radiat. Res. 10.1093/jrr/rrt090. 4. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adaptation. J. Gerontol. A Biol. Sci. Med. Sci. 10.1093/gerona/glt122. 5. Paneni, F. et al. (2013). Deletion of the activated protein-1 transcription factor JunD induces oxidative stress and accelerates age-related endothelial dysfunction. Circulation 127:1229-1240. 6. Connell, B.J. et al. (2012). UPEI-100, a conjugate of lipoic acid and apocynin, mediates neuroprotection in a rat model of ischemia/reperfusion. Am. J. Physiol. Regulatory Integrative Comp. Physiol. 302:R866-R895. 7. Zhang, Z. et al. (2012). TRPM2 Ca2+ channel regulates energy balance and glucose metabolism. Am. J. Phyisol. Endocrin. Metab. 302:E807-E816.

Standard Curve Using the OxiSelect™ Superoxide Dismutase Activity Assay. Product Name OxiSelect™ Superoxide Dismutase Activity Assay

Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-340

www.cellbiolabs.com

info@cellbiolabs.com

107

OXIDATIVE STRESS / DAMAGE

Antioxidant Assays

OxiSelect™ Total Glutathione (GSSG/GSH) Assay Kit Glutathione is an intracellular tripeptide thiol that protects cells from free radicals by acting as an antioxidant. Glutathione exists within cells in both reduced (GSH) and oxidized (GSSG) forms; it is involved in the breakdown of peroxides and also helps maintain exogenous antioxidants such as vitamins C and E. The OxiSelect™ Total Glutathione Assay Kit is a quantitative assay for measuring total combined GSH and GSSG content in a variety of sample types. Oxidized glutathione is enzymatically reduced, followed by colorimetric detection in a microplate reader.

 Sensitive: Detect as little as 8 nM total glutathione  Fast: Obtain results in less than 30 minutes  Versatile: Suitable for use with saliva, urine, serum, plasma, and cell or tissue lysates Recent Product Citations 1. Mani, S. et al. (2013). Decreased endogenous production of hydrogen sulfide accclerates atherosclerosis. Circulation 127:2523-2534. 2. Karakus, E. et al. (2013). Agomelatine: an antidepressant with new potent hepatoprotective effects on paracetamol-induced liver damage in rats. Human and Exp. Toxicol. 10.01177/0960327112472994.

Assay Principle for the OxiSelect™ Total Glutathione Assay Kit. In the presence of NADPH, glutathione reductase (provided) converts all glutathione into reduced form (GSH). The reduced glutathione then reacts with the provided chromogen to yield a color detectable at 405 nm. Product Name OxiSelect™ Total Glutathione (GSSG/GSH) Assay Kit

Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-312

OxiSelect™ Glutathione Reductase Assay Kit The OxiSelect™ Glutathione Reductase Assay Kit is a quantitative assay for measuring the activity levels of glutathione reductase in a variety of sample types. The assay principle is similar to that of our Total Glutathione Assay Kit above, except that endogenous levels of glutathione reductase drive the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH).  Sensitive: Detect activity levels as low as 0.6 mU/mL  Fast: Obtain results in less than 30 minutes  Versatile: Suitable for use with erythrocytes, plasma, cell lysates, or tissue extracts Product Name OxiSelect™ Glutathione Reductase Assay Kit

108

Phone 1-858-271-6500

Standard Curve Generated with the OxiSelect™ Glutathione Reductase Assay Kit. Various concentrations of glutathione reductase standard were tested according to the Assay Protocol. OD values were read at 1 minute increments at 405 nm. Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-812

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Antioxidant Assays

OXIDATIVE STRESS / DAMAGE

OxiSelect™ Cellular Antioxidant Assay Kit, for in vivo Evaluation of Exogenous Antioxidants Measuring the effects of antioxidant compounds in an in vitro assay may not accurately reflect their efficacy because such assays do not account for physiological conditions such as pH, temperature, uptake, metabolism, or the bioavailability or efficacy of an antioxidant compound. The OxiSelect™ Cellular Antioxidant Activity Assay Kit provides a mechanism to test exogenous antioxidants in a cell-based environment, delivering a more accurate measurement of the compound’s true physiological efficacy. A cell-permeable fluorometric dye is added to intact cells; when free radicals are generated, they bind to the dye producing a bright fluorescent signal. When the exogenous antioxidant is added, it eliminates the free radicals resulting in decreased fluorescence.

 Physiological: Measures the efficacy of an antioxidant compound in a cellular environment  More Accurate: Compared to in vitro assays that do not take into account pH, temperature, or other relevant intracellular conditions  Sensitive: Detects as little as 10 µM Quercetin, a bioflavonoid which serves as the assay’s standard  Fast: Obtain results in about one hour

18 16

Fluorescence (RFUs)

14 2000 uM Quercetin

1000 10

500

250 125

62.5 31.3

0 2 0 0

30 40 Time (Minutes)

Cellular Antioxidant Activity of Quercetin in HeLa Cells. 60,000 HeLa cells were seeded and cultured in a 96-well plate until confluent. Cells were then pretreated with DCFH-DA and Quercetin for 60 minutes at 37ºC. Free Radical Initiator was then added to the cells to begin the assay. Fluorescence readings were taken every 5 minutes for one hour at 37ºC.

Product Name OxiSelect™ Cellular Antioxidant Activity Assay Kit

Assay Principle for the OxiSelect™ Cellular Antioxidant Activity Assay Kit. An exogenous antioxidant compound is added to cells along with DCFH-DA dye. Upon entry into the cell, the DCFHDA is cleaved to DCFH which can bind reactive oxygen species (ROS) generated within the cell by the addition of a free radical initiator. Binding of DCFH to ROS yields DCF which produces a bright fluorescence. The presence of the exogenous antioxidant compound reduces the ROS available to the DCFH dye, yielding a lower fluorescent signal.

Detection

Size

Catalog Number

Fluorometric

192 Assays

STA-349

www.cellbiolabs.com

info@cellbiolabs.com

109

OXIDATIVE STRESS / DAMAGE

Antioxidant Assays

OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit The OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit measures the total antioxidant capacity of biomolecules from a variety of sample types via a Single Electron Transfer (SET) mechanism. The assay works with a variety of antioxidants and is suitable for testing plasma, serum, urine, cell lysates, tissue homogenates and food extracts.

OxiSelect™ Total Antioxidant Capacity Assay Principle.

Recent Product Citations 1. Stark, M. et al. (2013). Differential effects of docosahexaenoic acid on preterm and term placental pro-oxidant/antioxidant balance. Reproduction 146:243-251. 2. Bakalova, R. et al. (2013). Tissue redox activity as a hallmark of carcinogenesis: from early to terminal stages of cancer. Clin. Cancer Res. 19:2503-2517. 3. Wang, Y. et al. (2013). Therapeutic effect of MG-132 on diabetic cardiomyopathy is associated with its suppression of proteasomal activities: roles of Nrf2 and NF-kB. Am. J. Physiol. Heart Circ. Physiol. 304:H567-H578. Product Name OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit

Detection

Size

Catalog Number

Colorimetric

200 Assays

STA-360

OxiSelect™ ORAC and HORAC Activity Assay Kits The ORAC (Oxygen Radical Antioxidant Capacity) and HORAC (Hydroxyl Radical Antioxidant Capacity) assays measure the antioxidant capacity of biomolecules against peroxyl radicals and hydroxyl radicals, respectively. The assays are suitable for plasma, cell fractions, and tissue lysates, as well as solid and aqueous nutrition samples.

Recent Product Citations 1. Ungvari, Z. et al. (2013). Testing predictions of the oxidative stress hypothesis of aging using a novel invertebrate model of longevity: the giant clam (Tridacna derasa). J. Gerontol. A. Biol. Sci. Med. Sci. 68:359-367. 2. Bailey-Downs, L.C. et al. (2011). Liver-specific knockdown of IGF-1 decreases vascular oxidative stress resistance by impairing the Nrf2-dependent antioxidant response: a novel model of vascular aging. J. Gerontol. A. Biol. Sci. Med. Sci. 10.1093/ gerona/glr164.

Assay Principle for the OxiSelect™ Oxygen Radical Antioxidant Capacity (ORAC) Assay. Product Name

110

Detection

OxiSelect™ ORAC Activity Assay Kit

Fluorometric

OxiSelect™ HORAC Activity Assay Kit

Fluorometric

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

192 Assays

STA-345

5 x 192 Assays

STA-345-5

192 Assays

STA-346

5 x 192 Assays

STA-346-5

Fax 1-858-271-6514

CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein Small GTPase / G-Protein Signaling Small GTP-binding proteins (GTPases) regulate a variety of cell signaling pathways and are therefore involved in a wide range of cell functions, processes, and morphology. The most studied small GTPases include Ras, Rac, Rho and Cdc42. We offer a variety of tools to enable the study of these small GTPase family members:    

Small GTPase Activation Assays Small GTPase Activation ELISA Kits Small GTPase Assay Beads Active Rac-GEF Assay

   

Small GTPase Expression Vectors Small GTPase Premade Adenoviruses Small GTPase Retroviral Constructs Small GTPase Recombinant Proteins

In addition, we offer sensitive assays to detect cyclic AMP and cyclic GMP, both of which are important regulators in the G-Protein signaling cascade.

Small GTPase Activation Assays Our Small GTPase Activation Assays use visible agarose beads to selectively pull down the active form of the target of interest. The precipitated GTPase is then detected by Western blot using a target specific antibody included in the kit.

 Safe: Non-radioactive assay format  Visual Check: Agarose beads can be easily seen  Fast Results: 1 hour plus electrophoresis/blotting

If you are studying more than one small GTPase target, consider one of our Small GTPase Activation Assay Combo Kits. These combo kits allow you to measure the following targets at a savings compared to buying separate kits for each target:  

Rac1 and Cdc42 RhoA, Rac1 and Cdc42

Visible Agarose Beads Provided in the Small GTPase Activation Assays. Beads are easy to visualize, making it easier to avoid potential loss during washes and aspirations.

Small GTPase Activation Assay Principle.

112

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Small GTPase/G-Protein CELL SIGNALING & PROTEIN BIOLOGY Small GTPase Activation Assays (continued) Recent Product Citations 1. Ohashi, K. et al. (2012). Lung cancers with acquired resistance to EGFR inhibitors occasionally harbor BRAF gene mutations but lack mutations in KRAS, NRAS or MEK1. PNAS 109:E2127-E2133. (STA-400) 2. Geryk-Hall, M. et al. (2010). Driven to death: inhibition of farnesylation increases Ras activity in osteosarcoma and promotes growth arrest and cell death. Mol. Cancer Ther. 9:1111-1119. (STA-400) 3. Camalier, C.E. et al. (2010). Elevated phosphate activates N-ras and promotes cell transformation and skin tumorigenesis. Cancer Prev. Res. 3:359-370. (STA-400) 4. Tsukamoto, Y. et al. (2013). A novel heart failure mice model of hypertensive heart disease by angiotensin II infusion, nephrectomy, and salt loading. Am. J. Physiol. Heart Circ. Physiol. 305:H1658-H1667. (STA-401-1) 5. Petzold, T. et al. (2013). ß1 integrin-mediated signals are required for platelet granule secretion and hemostasis in mouse. Blood 122:27232731. (STA-401-1) 6. Cristante, E. et al. (2013). Identification of an essential endogenous regulator of blood-brain barrier integrity, and its pathological and therapeutic implications. PNAS 110:832-841. (STA-401-1) 7. Yanagashita, T. et al. (2014). Actin-binding protein, espin: a novel metastatic regulator for melanoma. Mol. Cancer Res. 12:440-446. (STA401-1, STA-403-A) 8. He, S. et al. (2013). SRGAP1 is a candidate gene for papillary thyroid carcinoma susceptibility. J. Clin. Endocrinol. Metab. 98:E973-E980. (STA-402) 9. Fiuza, M. et al. (2013). GluN3A expression restricts spine maturation via inhibition of GIT1/Rac1 signaling. PNAS 110:20807-20812. (STA403-A) 10. Basu, M. et al. (2012). Wnt/ß-catenin pathway is regulated by PITX2 homeodomain protein and thus contributes to the proliferation of human ovarian adenocarcinoma cell SKOV-3. J. Biol. Chem. 288:4355-4367. (STA-403-A) 11. Holmes, K.M. et al. (2012). Insulin-like growth factor-binding protein 2-driven glioma progression is prevented by blocking a clinically significant integrin, integrin-linked kinase, and NF-kB network. PNAS 109:2168-2173. (STA-404) 12. Wang, J. et al. (2013). DEK depletion negatively regulates Rho/ROCK/MLC pathway in non-small cell lung cancer. J. Histochem. & Cytochem. 10.1369/00221155413488120. (STA-405) 13. Quint, P. et al. (2013). Sphingosine 1-phosphate (S1P) receptors 1 and 2 coordinately induce mesenchymal cell migration through S1P activation of complementary kinase pathways. J. Biol. Chem. 288:5398-5406. (STA-405) 14. Baranwai, S. et al. (2011). Molecular characterization of the tumor-suppressive function of nischarin in breast cancer. J. Natl. Cancer Inst. 10.1093/jnci/djr350. (STA-405) 15. Grossman, A. et al. (2013). The small GTPase ARF6 stimulates ß-catenin transcriptional activity during WNT5A-mediated melanoma invasion and metastasis. Sci Signal 6:ra14. (STA-407-6) Product Name

Detection

Size

Catalog Number

Arf1 Activation Assay

Immunoblot/ECL

20 Assays

STA-407-1

Arf6 Activation Assay

Immunoblot/ECL

20 Assays

STA-407-6

Cdc42 Activation Assay

Immunoblot/ECL

20 Assays

STA-402

Rac1 Activation Assay

Immunoblot/ECL

20 Assays

STA-401-1

Rac2 Activation Assay

Immunoblot/ECL

20 Assays

STA-401-2

Ral Activation Assay

Immunoblot/ECL

20 Assays

STA-408

Ran Activation Assay

Immunoblot/ECL

20 Assays

STA-409

Rap1 Activation Assay

Immunoblot/ECL

20 Assays

STA-406-1

Rap2 Activation Assay

Immunoblot/ECL

20 Assays

STA-406-2

Pan-Ras Activation Assay

Immunoblot/ECL

20 Assays

STA-400

H-Ras Activation Assay

Immunoblot/ECL

20 Assays

STA-400-H

K-Ras Activation Assay

Immunoblot/ECL

20 Assays

STA-400-K

N-Ras Activation Assay

Immunoblot/ECL

20 Assays

STA-400-N

RhoA Activation Assay

Immunoblot/ECL

20 Assays

STA-403-A

RhoB Activation Assay

Immunoblot/ECL

20 Assays

STA-403-B

RhoC Activation Assay

Immunoblot/ECL

20 Assays

STA-403-C

Rac1/Cdc42 Activation Assay Combo Kit

Immunoblot/ECL

20 Assays/Target

STA-404

RhoA/Rac1/Cdc42 Activation Assay Combo Kit

Immunoblot/ECL

10 Assays/Target

STA-405

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113

CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein 96-Well Ras Activation ELISA Kits Our 96-Well Ras Activation Assays use the Raf1 Rho binding domain (Raf1 RBD) to selectively pull down the active form of Ras from purified or endogenous samples. The captured GTP-Ras is then detected by a pan-Ras antibody and HRP-conjugated secondary antibody. Detection is by either colorimetric or chemiluminescent plate reader.

Pan-Ras Antibody Specificity. Anti-pan-Ras antibody reactivity with H-Ras, K-Ras and N-Ras human isoforms by dot blot. EGF Stimulation and Active Ras Detection with the 96-Well Ras Activation ELISA Kit. HeLa cells were serum starved for 18 hours before EGF stimulation of 50 ng/mL for 2 minutes. Lysates were then prepared according to the assay protocol.

Product Name 96-Well Ras Activation ELISA Kit

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-440

Chemiluminescent

96 Assays

STA-441

Active Rac-GEF Assay Kit (Tiam1) Guanine nucleotide exchange factors (GEFs) activate small GTPases by catalyzing the exchange of GDP for GTP. Tiam1 Activation Assay. Left: 293 cells were transfected with active Tiam1. Active Tiam1 in lysate was pulled down with Rac1 G15A agarose beads. Right: Active Tiam-1 in 2 mg of MDA-231 cell lysate was pulled down with Rac1 G15A agarose beads and probed with anti-Tiam1 antibody.

Our Active Rac-GEF Assay Kit (Tiam1) uses the agarose bead technology of our Small GTPase Activation Assays (previous page). Agarose beads pull down the active form of Rac-GEFs from endogenous lysates or purified samples. The specific GEF known as Tiam1 is then specifically detected with a polyclonal antibody.

Recent Product Citation Oubaha, M. et al. (2012). Formation of a PKC/Ă&#x;-catenin complex in endothelial cells promotes angiopoietin-1-induced collective directional migration and angiogenic sprouting. Blood 120:3371-3381.

Product Name Active Rac-GEF Assay Kit (Tiam1)

114

Phone 1-858-271-6500

Detection

Size

Catalog Number

Immunoblot/ECL

20 Assays

STA-422

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Small GTPase/G-Protein CELL SIGNALING & PROTEIN BIOLOGY Small GTPase Agarose Assay Beads Our agarose beads are useful for selectively pulling down only the active form of small GTPases. The beads are colored for easily visualization. These are the same beads used in our Small GTPase Activation Assays (p. 108-109).

Visible Agarose Beads. Beads are easy to visualize, making it easier to avoid potential loss during washes and aspirations.

Product Name

Recent Product Citations 1. Yuen, H. et al. (2013). RanGTPase: a candidate for Mycmediated cancer progression. J. Natl. Cancer Inst. 105:475-488. (STA-410) 2. Moniz, S. et al. (2007). Protein kinase WNK2 inhibits cell proliferation by negatively modulating the activation of MEK1/ERK1/2. Oncogene 26(41):6071-6081. (STA-410) 3. Pothula, S. et al. (2013). Regulation of Cdc42 expression and signaling is critical for promoting corneal epithelial wound healing. Invest. Ophthalmol. Vis. Sci. 54:5343-5352. (STA-411) 4. Zhang, Q-G. et al. (2009). Estrogen attenuates ischemic oxidative damage via an estrogen receptor alpha-mediated inhibition of NADPH oxidase activation. J. Neurosci. 29:13823-13836. (STA-411) 5. Levy-Adam, F. et al. (2008). Heparanase facilitates cell adhesion and spreading by clustering of cell surface heparan sulfate proteoglycans. PLoS ONE 3(6):e2319. (STA-411, STA-412) 6. Sabbatini, M. E. et al. (2010). CCK activates RhoA and Rac1 differentially through G-alpha-13 and G-alpha-q in mouse pancreatic acini. Am. J. Physiol. Cell Physiol. 298:C592-C605. (STA -411, STA-412) 7. Sabbatini, M. et al. (2008). Rap1 activation plays a regulatory role in pancreatic amylase secretion. J. Biol. Chem. 283:2388423894. (STA-412)

Target

Size

Catalog Number

GGA3 PBD Agarose Beads

Arf

400 µg

STA-419

PAK1 PBD Agarose Beads

Cdc42, Rac

400 µg

STA-411

Raf1 RBD Agarose Beads

Ras

400 µg

STA-410

RalBP1 PBD Agarose Beads

Ral

400 µg

STA-420

RalGDS RBD Agarose Beads

Rap

400 µg

STA-418

RanBP1 Agarose Beads

Ran

400 µg

STA-421

Rhotekin RBD Agarose Beads

Rho

400 µg

STA-412

GEF Agarose Assay Beads Our GEF agarose beads are useful for selectively pulling down only the active form of guanine nucleotide exchange factors (GEF). The beads are colored for easily visualization.

Product Name

Recent Product Citations 1. Ngok, S. et al. (2013). Phosphorylation-mediated 14-3-3 protein binding regulates the function of the Rho-specific guanine nucleotide exchange factor (RhoGEF) Syx. J. Biol. Chem. 288:6640-6650. (STA-431) 2. Colacios, C. et al. (2011). The p.Arg63Trp polymorphism controls Vav1 functions and Fox3p regulatory T cell development. J. Exp. Med. 208:2183-2191. (STA-432) Target

Size

Catalog Number

Cdc42 G15A Agarose Beads

Cdc42-GEF

800 µg

STA-433

Rac1 G15A Agarose Beads

Rac1-GEF

800 µg

STA-432

RhoA G17A Agarose Beads

RhoA-GEF

400 µg

STA-431

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115

CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein Small GTPase Expression Vector Sets Our Small GTPase Expression Vectors are ideal tools for the study of the most commonly researched small GTPase targets. Each set contains 3 mammalian expression vectors:   

Wild type Dominant negative Constitutively active

Vectors are available with or without a GFP reporter gene. All vectors are supplied as bacterial glycerol stocks.

Product Name

Vectors

Size

Catalog Number

Cdc42 Expression Vector Set

Wild Type, T17N (Dom. Neg.), Q61L (Active)

3 x 100 µL

STA-455

GFP-Cdc42 Expression Vector Set

Wild Type, T17N (Dom. Neg.), Q61L (Active)

3 x 100 µL

STA-451

Rac1 Expression Vector Set

Wild Type, T17N (Dom. Neg.), G12V (Active)

3 x 100 µL

STA-454

GFP-Rac1 Expression Vector Set

Wild Type, T17N (Dom. Neg.), Q61L (Active)

3 x 100 µL

STA-450

H-Ras Expression Vector Set

Wild Type, T17N (Dom. Neg.), G12V (Active)

3 x 100 µL

STA-457

RhoA Expression Vector Set

Wild Type, T19N (Dom. Neg.), G14V (Active)

3 x 100 µL

STA-456

GFP-RhoA Expression Vector Set

Wild Type, T19N (Dom. Neg.), Q63L (Active)

3 x 100 µL

STA-452

Active Small GTPase Expression Vector Sets Our Active Small GTPase Expression Vectors are similar to the expression vectors above, except that they are provided as sets of 3 different active mutants for a single small GTPase target. All vectors are supplied as bacterial glycerol stocks. Product Name

Vectors

Size

Catalog Number

Active Rac1 Expression Vector Set

Q61L, Q61L/F37A, Q61L/Y40C

3 x 100 µL

STA-458

Active H-Ras Expression Vector Set

V12, V12/S35, V12/C40

3 x 100 µL

STA-459

Size

Catalog Number

3 x 100 µL

STA-460

Exoenzyme C3 (Rho Inhibitor) Expression Vector This vector is supplied as bacterial glycerol stock. Product Name Exoenzyme C3 Expression Vector

116

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Small GTPase/G-Protein CELL SIGNALING & PROTEIN BIOLOGY Gene-Specific Recombinant Retroviral Vectors Our recombinant retroviral plasmids contain a specific gene cloned into a pBABE vector backbone. Each vector is supplied as 100 µL of bacterial glycerol stock.

Recent Product Citation Zhao, B. et al. (2012). TNF-induced osteoclastogenesis and inflammatory bone resorption are inhibited by transcription factor RBP-J. J. Exp. Med. 209:319-334. (RTV-101)

Target Name

Vector Backbone

Catalog Number

Vector Backbone

Catalog Number

Cdc42 L61

pBABEhygro

RTV-203

N-Ras K61

pBABEpuro

RTV-222

myr-Rac1

pBABEpuro

RTV-201

Ras V12

pBABEpuro

RTV-101

Rac1 V12

pBABEhygro

RTV-202

Ras V12C40

pBABEpuro

RTV-104

Rac3 V12

pBABEhygro

RTV-205

Ras V12G37

pBABEpuro

RTV-103

K-Ras

pBABEpuro

RTV-220

Ras V12S35

pBABEpuro

RTV-102

K-Ras Q61

pWZLhygro

RTV-221

RhoA L63

pBABEhygro

RTV-204

Small GTPase Recombinant Adenoviruses All of Cell Biolabs’ premade recombinant adenoviruses contain 5 x 109 viral particles per vial. They are provided as 50 µL aliquots at a concentration of 1 x 1011 viral particles/mL in TBS with 10% glycerol. Recent Product Citations 1. Black, S.A. et al. (2008). TGFß1 stimulates connective tissue growth factor (CCN2/CTGF) expression in human gingival fibroblasts through a RhoA-independent, Rac1/Cdc42dependent mechanism: statins with forskolin block TGFß1induced CCN2/CTGF expression. J. Biol. Chem. 283:1083510847. (ADV-145, ADV-150, ADV-153, ADV-156) 2. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires avß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Biol. Cell 23:1104-1114. (ADV-150) 3. Thomas, M.A. et al. (2009). E4orf1 limits the oncolytic potential of the E1B-55K-deleted adenovirus. J. Virol. 83:2406-2416. (ADV-150) 4. Rendon, B. et al. (2007). Regulation of human lung adenocarcinoma cell migration and invasion by MIF. J. Biol. Chem. 282:29910-29918. (ADV-150) 5. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150, ADV-153, ADV-154) 6. Cheng, Z.-J. et al. (2010). Co-regulation of caveolar and Cdc42 -dependent fluid phase endocytosis by phosphocaveolin-1. J. Biol. Chem. 285:15119-15125. (ADV-153) 7. Perez-Moreno, M. et al. (2008). Loss of p120 catenin and links to mitotic alterations, inflammation, and skin cancer. PNAS 105:15399-15404. (ADV-156) 8. Neal, M. et al. (2013). A critical role for TLR4 induction of autophagy in the regulation of enterocyte migration and the pathogenesis of necrotizing enterocolitis. J. Immunol. 190:35413551. (ADV-156, ADV-157) 9. Vaught, D. et al. (2009). Regulation of mammary gland branching morphogenesis by EphA2 receptor tyrosine kinase. Mol. Biol. Cell 20:2572-2581. (ADV-157) 10.Brantley-Sieders, D. et al. (2008). The receptor tyrosine kinase EphA2 promotes mammary adenocarcinoma tumorigenesis and metastatic progression in mice by amplifying ErbB2 signal. J. Clin. Invest. 118:64-78. (ADV-157)

Target Name

Catalog Number

Cdc42

ADV-152

Cdc42 L61 (Constitutively Active)

ADV-154

Cdc42 N17 (Dominant Negative)

ADV-153

Rac1

ADV-149

Rac1 L61 (Constitutively Active)

ADV-151

Rac1 N17 (Dominant Negative)

ADV-150

Ras N17 (Dominant Negative)

ADV-145

Ras V12 (Constitutively Active)

ADV-146

Ras V12C40

ADV-148

Ras V12S35

ADV-147

Rho L63 (Constitutively Active)

ADV-157

Rho N19 (Dominant Negative)

ADV-156

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117

CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein Small GTPase Recombinant Human Proteins Protein Name

Tag / Location

Size

Catalog Number

Protein Name

Tag / Location

Size

Catalog Number

Cdc42

T7 / N-term

50 µg

STA-700

Rab 35

6xHis / N-term

10 µg

STA-726

Rab1a

6xHis / N-term

10 µg

STA-701

Rab IF

6xHis / N-term

10 µg

STA-727

Rab1b

6xHis / N-term

10 µg

STA-702

Rac1

6xHis / N-term

50 µg

STA-728

Rab2a

6xHis / N-term

20 µg

STA-703

Rac1

None

50 µg

STA-729

Rab3a

6xHis / N-term

20 µg

STA-704

Rac2

6xHis / N-term

20 µg

STA-730

Rab3b

6xHis / N-term

20 µg

STA-705

Rac3

None

20 µg

STA-731

Rab3d

6xHis / N-term

20 µg

STA-706

Ral A

6xHis / N-term

25 µg

STA-732

Rab4a

6xHis / N-term

10 µg

STA-707

Ral B

6xHis / N-term

10 µg

STA-733

Rab5a

None

20 µg

STA-708

Ran

6xHis / N-term

10 µg

STA-734

Rab5b

6xHis / N-term

20 µg

STA-709

Rap1a

6xHis / N-term

10 µg

STA-735

Rab6a

6xHis / N-term

10 µg

STA-710

Rap1b

6xHis / N-term

10 µg

STA-736

Rab6b

6xHis / N-term

20 µg

STA-711

Rap2a

6xHis / N-term

10 µg

STA-737

Rab7a

6xHis / N-term

20 µg

STA-712

H-Ras

6xHis / C-term

25 µg

STA-747

Rab11a

6xHis / N-term

25 µg

STA-713

K-Ras

6xHis / N-term

25 µg

STA-748

Rab13

6xHis / N-term

10 µg

STA-714

N-Ras

None

10 µg

STA-749

Rab14

6xHis / N-term

10 µg

STA-715

R-Ras

6xHis / N-term

10 µg

STA-750

Rab17

6xHis / N-term

20 µg

STA-716

R-Ras2

6xHis / N-term

50 µg

STA-751

Rab18

T7 / N-term

10 µg

STA-717

RERG

6xHis / N-term

10 µg

STA-738

Rab21

6xHis / N-term

10 µg

STA-718

RHEB

T7 / N-term

50 µg

STA-739

Rab22

6xHis / N-term

5 µg

STA-719

RhoA

6xHis / N-term

20 µg

STA-740

Rab23

6xHis / N-term

20 µg

STA-720

RhoB

6xHis / N-term

10 µg

STA-741

Rab24

6xHis / N-term

20 µg

STA-721

RhoC

6xHis / N-term

10 µg

STA-742

Rab27a

6xHis / N-term

25 µg

STA-722

RhoD

6xHis / N-term

5 µg

STA-743

Rab27b

6xHis / N-term

20 µg

STA-723

RND1

6xHis / N-term

20 µg

STA-744

Rab32

6xHis / N-term

20 µg

STA-724

RND3

6xHis / N-term

20 µg

STA-745

Rab34

6xHis / N-term

10 µg

STA-725

SAR1A

6xHis / N-term

20 µg

STA-746

Regulators of G Protein Signaling (RGS) Recombinant Human Proteins

118

Protein Name

Tag / Location

Size

Catalog Number

Protein Name

Tag / Location

Size

Catalog Number

RGS2

6xHis / N-term

5 µg

STA-780

RGS16

6xHis / N-term

10 µg

STA-784

RGS4

6xHis / N-term

10 µg

STA-781

RGS17

6xHis / N-term

25 µg

STA-785

RGS5

6xHis / N-term

25 µg

STA-782

RGS19

6xHis / N-term

20 µg

STA-786

RGS10

6xHis / N-term

25 µg

STA-783

RGS21

6xHis / N-term

20 µg

STA-787

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Small GTPase/G-Protein CELL SIGNALING & PROTEIN BIOLOGY Cyclic AMP and GMP ELISA Kits

Recent Product Citations 1. Meyer, R. et al. (2013). GPR37 and GPR37L1 are receptors for the neuroprotective and glioprotective factors prosaptide and prosaposin. PNAS 110:9529-9534. (STA-500) 2. Sun, Z. et al. (2011). The WD40 repeat protein WDR26 binds Gßg and promotes Gßg-dependent signal transduction and leukocyte migration. J. Biol. Chem. 286:43902-43912. (STA500) 3. Chen, M. et al. (2010). Involvement of cAMP in nerve growth factor-triggered p35/Cdk5 activation and differentiation in PC12 cells. Am. J. Physiol. Cell Physiol. 299:C516-C527. (STA-500) 4. McCoy, K.L. et al. (2010). PAR1 and PAR2 couple to overlapping and distinct sets of G proteins and linked signaling pathways to differentially regulate cell physiology. Mol. Pharmacol. 77:1005-1015. (STA-500) 5. Liu, L. et al. (2012). Radil controls neutrophil adhesion and motility through ß2-integrin activation. Mol. Biol. Cell 23:4751-4765. (STA-501)

 Sensitive: Detect as little as 1 pmol/mL  Versatile: Suitable for use with cell and tissue lysates, urine, plasma, or culture medium  Convenient: Strip-well plate format with either colorimetric or chemiluminescent detection

2.5

OD 450nm

Cyclic AMP and cyclic GMP are important regulatory molecules in the GPCR signaling cascade. Our cAMP and cGMP ELISA Kits provide a highly sensitive method to measure low levels of cAMP or cGMP in a variety of sample types.

1.5

0.5

0 0.0

0.1

1.0

10.0

100.0

1000.0

10000.0 100000.0

cAMP (pmol/mL)

Standard Curve Created with the cAMP ELISA Kit, Colorimetric Format.

Product Name

Detection Colorimetric

cAMP ELISA Kit Chemiluminescent

Colorimetric cGMP ELISA Kit Chemiluminescent

Size

Catalog Number

96 Assays

STA-500

5 x 96 Assays

STA-500-5

96 Assays

STA-501

5 x 96 Assays

STA-501-5

96 Assays

STA-505

5 x 96 Assays

STA-505-5

96 Assays

STA-506

5 x 96 Assays

STA-506-5

Recombinant GRP-PH Domain The GRP1 (general receptor for phosphoinositide) protein binds phosphatidylinositol-3,4,5-triphosphate (PIP3) through a pleckstrin homology (PH) domain and displays a region of high sequence similarity to the yeast Sec7 protein. This recombinant protein is expressed and purified from E. coli as a fusion protein, and is provided at 1.0 mg/ ml in 1X PBS. Product Name Recombinant GRP-PH Domain

www.cellbiolabs.com

Size

Catalog Number

100 µg

STA-200

1 mg

STA-200-1MG

info@cellbiolabs.com

119

CELL SIGNALING & PROTEIN BIOLOGY Kinase Assays Rho Kinase (ROCK) Activity Assays Rho Kinase (ROCK) is a serine/threonine kinase which is a target of Rho. ROCK mediates Rho signaling and reorganizes the actin cytoskeleton via the phosphorylation of several substrates that contribute to contractility and the assembly of actin filaments. Our ROCK Activity Assays provide a non-radioactive format to measure the level of active ROCK in cell or tissue lysates. The immunoblot kit provides a convenient format for measuring ROCK activity in a few samples, while the 96-well Activity Assay contains a stripwell plate precoated with MYPT1 for higher throughput.

Results Using the ROCK Activity Immunoblot Kit. Lanes 1, 3, 5, 7: Without ROCK (negative control). Lanes 2, 4, 6, 8: With ROCK. Lanes 1 & 2: 200 ng MYPT1; Lanes 3 & 4: 100 ng; Lanes 5 & 6: 50 ng; Lanes 7 & 8: 25 ng. Phosphorylation of MYPT1 substrate was detected by anti-phospho-MYPT1 as described in the protocol. Recent Product Citations 1. Georgess, D. et al. (2014). Comparative transcriptomics reveals RhoE as a novel regulator of actin dynamics in bone-resorbing osteoclasts. Mol. Biol. 25:380-396. (STA-415) 2. Wang, J.N. et al. (2011). Response gene to complement 32 promotes vascular lesion formation through stimulation of smooth muscle cell proliferation and migration. Arterioscler. Thromb. Vasc. Biol. 31:e19-e26. (STA-415) 3. Xiao, L. et al. (2009). ROCK mediates phorbol ester-induced apoptosis in prostate cancer cells via p21-Cip1 upregulation and JNK. J. Biol. Chem. 284:29365-29375. (STA-415) 4. Burger, D. et al. (2011). Endothelial microparticle formation by angiotensin II is mediated via ang II receptor type I/NADPH oxidase/Rho kinase pathways targeted to lipid rafts. Arterioscler. Thromb. Vasc. Biol. 31:1898-1907. (STA-416) 5. Haas, B. et al. (2009). Protein kinase G controls brown fat cell differentiation and mitochondrial biogenesis. Sci. Signal. 2:ra78. (STA-416) Product Name

120

96-Well ROCK Activity Assay Principle. Detection

Size

Catalog Number

ROCK Activity Immunoblot Kit

Immunoblot

20 Assays

STA-415

96 Assays

STA-416

96-Well ROCK Activity Assay

Colorimetric

5 x 96 Assays

STA-416-5

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Kinase Assays CELL SIGNALING & PROTEIN BIOLOGY Checkpoint Kinase Activity Assays Checkpoint kinases, including CHK1 and CHK2, can be activated in response to DNA damage prior to mitosis. These kinases phosphorylate Cdc25C, a protein phosphatase, at Ser-216. This phosphorylation ultimately leads to cell cycle arrest, preventing mitosis and avoiding the passage of DNA damage to daughter cells. Our Checkpoint Kinase Activity Assays allow you to conveniently measure the activity of CHK1 and CHK2. The assays use recombinant Cdc25C as a checkpoint kinase substrate. Phosphorylated Cdc25C (Ser216) is detected using a phosphospecific antibody. Checkpoint Kinase Activitiy Assays are available in two formats: a Western blot assay and a 96-well plate-based activity assay.

CHK1 Activity Using the Checkpoint Kinase Activity Immunoblot Kit. Lane 1: Negative Control; Lane 2: 10 ng of active CHK1. Product Name

96-Well Checkpoint Kinase Activity Assay Principle. Detection

Size

Catalog Number

Checkpoint Kinase Activity Immunoblot Kit

Immunoblot

20 Assays

STA-413

96 Assays

STA-414

96-Well Checkpoint Kinase Activity Assay Kit

Colorimetric

5 x 96 Assays

STA-414-5

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info@cellbiolabs.com

121

CELL SIGNALING & PROTEIN BIOLOGY Reporter Molecules Reporter Assays, Cell Lines and Reagents We offer a variety of tools for various reporter molecules:   

Reporter Assays Reporter Stable Cell Lines Recombinant Fluorescent Proteins

 

Antibodies to Reporter Molecules Reporter Viral Vectors

GFP ELISA Kit Most imaging studies of rGFP are qualitative, and quantitation by FACS is time consuming and expensive. Our GFP ELISA kit uses a standard microplate reader to quantify GFP levels with extremely high sensitivity. This kit will detect GFP form Aequorea victoria as well as its variants.

Standard Curve Generated with the GFP ELISA Kit.

Product Name GFP ELISA Kit

 Sensitive: Detection limit of 30 pg/mL  Versatile: Quantify GFP and its variants: BFP, CFP or YFP  Easy Quantitation: Measure GFP levels in a standard microplate reader Recent Product Citations 1. Mango, R. et al. (2014). C-C chemokine receptor 5 on pulmonary mesenchymal cells promotes experimental metastasis via the induction of erythroid differentiation regulator 1. Mol. Cancer Res. 12:274-282. 2. Mitchell, A. et al. (2014). Promyelocytic leukemia protein is a cell -intrinsic factor inhibiting parvovirus DNA replication. J. Virol. 88:925-936. 3. Coghill, J.M. et al. (2013). CC chemokine receptor 8 potentiates donor Treg survival and is critical for the prevention of murine graft-versus-host disease. Blood 122:825-836. 4. Pedersen, J. et al. (2012). Glucose metabolism is altered after loss of L cells and -cells but not influenced by loss of K cells. Am. J. Physiol. Endocrinol. Metab. 304:E60-E73. 5. Zhou, W. et al. (2012). Inducible podocyte injury and proteinuria in transgenic Zebrafish. J. Am. Soc. Nephrol. 23:1039-1047. 6. Fonseca, J.P. et al. (2012). In vivo polycomb kinetics and mitotic chromatin binding distinguish stem cells from differentiated cells. Genes and Dev. 26: 857-871. 7. Coghill, J.M. et al. (2010). Separation of graft-versus-host disease from graft-versus-leukemia responses by targeting CCchemokine receptor 7 on donor T cells. Blood 115:4914-4922. 8. Rajan, S. et al. (2010). In vitro processing and secretion of mutant insulin proteins that cause permanent neonatal diabetes.

Detection

Size

Catalog Number

Colorimetric

96 Assays

AKR-121

GFP Quantitation Kit When direct quantitation of GFP fluorescence levels is desired, our GFP Quantitation Kit provides a superior method over time-consuming flow cytometry and semi-quantitative imaging techniques. This kit measures fluorescence levels directly in a platebased fluorometer. Product Name GFP Quantitation Kit

122

Phone 1-858-271-6500

Recent Product Citations 1. Pfeiffer, B. et al. (2012). Using translational enhancers to increase transgene expression in Drosophila. PNAS 109:66266631. 2. Wamboldt, Y. et al. (2009). Participation of leaky ribosome scanning in protein dual targeting by alternative translation initiation in higher plants. Plant Cell 21:157-167.

Detection

Size

Catalog Number

Fluorometric

100 Assays

AKR-120

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Reporter Molecules CELL SIGNALING & PROTEIN BIOLOGY RFP ELISA Kit Our RFP ELISA Kit provides a convenient, sensitive alternative to imaging systems and time-consuming FACS quantitation. The assay quantifies a wide variety of red fluorescent protein variants including DsRed, TagRFP, TurboRFP, tdTomato, mCherry, mKate, mRuby, mBanana, mOrange, mPlum, and mStrawberry.

 Sensitive: Detection limit of 150 pg/mL  Easy Quantification: Measure RFP levels in a standard 96well microplate reader

Product Name

Detection

Size

Catalog Number

RFP ELISA Kit

Colorimetric

96 Assays

AKR-122

ß-Galactosidase Staining Kit LacZ is a commonly used reporter gene in transfection experiments because its gene product, ßgalactosidase, is extremely stable and resistant to proteolytic degradation, making it easy to assay. Our ß-Galactosidase Staining Kit provides an efficient, easy-to-use method to determine the transfection efficiency of the LacZ gene. Recent Product Citation Black, S.A. et al. (2008). TGFß1 stimulates connective tissue growth factor (CCN2/CTGF) expression in human gingival fibroblasts through a RhoA-independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGFß1-induced CCN2/ CTGF expression. J. Biol. Chem. 283:10835-10847.

X-Gal Staining of Infected HUVEC Cells and Chick CAM Tissue. Left: HUVEC cells were infected with purified Ad-ß-Gal at 50 MOI (multiplicity of infection). X-gal staining was performed after 48 hour infection period. Right: Purified Ad-ß-Gal was injected intravenously into a 10-day old chick embryo. After three days, X-gal staining was performed on the chick chorioallanoic membrane tissue.

Product Name ß-Galactosidase Staining Kit

Size

Catalog Number

75 Assays

AKR-100

Reporter Stable Cell Lines Each cell line expresses one or more reporter molecules. Recent Product Citations 1. Zhang, K. et al. (2014). Block-cellprinting for live single-cell printing. PNAS 111:2948-2953. (AKR-201, AKR211, AKR-252) 2. Zhang, W. et al. (2012). Microfluidics separation reveals the stem-cell-like deformability of tumor-initiating cells. PNAS 109:18707-18712. (AKR-211, AKR-252) 3. Weerakkody, D. et al. (2013). Family of pH (low) insertion peptides for tumor targeting. PNAS 110:5834-5839. (AKR213)

Cell Line

Catalog Number

293/YFP

AKR-280

MDA-MB-231/RFP

AKR-251

293T/GFP-Puro

AKR-202

MDA-MB-436/GFP

AKR-203

A549/GFP

AKR-209

MDA-MB-436/RFP

AKR-252

BT-549/GFP

AKR-255

MDA-MB-468/GFP

AKR-204

ES-2/GFP

AKR-206

NIH3T3/GFP

AKR-214

HeLa/GFP

AKR-213

OVCA429/GFP

AKR-212

MCF-7/GFP

AKR-211

OVCAR-5/RFP

AKR-254

Cell Line

Catalog Number

MCF-7/Luc

AKR-234

SKOV-3/GFP-Luc

AKR-225

293/CFP

AKR-270

MDA-MB-231/GFP

AKR-201

SKOV-3/Luc

AKR-232

293/GFP

AKR-200

MDA-MB-231/GFP-RFP

AKR-221

SKOV-3/RFP

AKR-253

293/Luc

AKR-230

MDA-MB-231/Luc

AKR-231

T47D/GFP

AKR-208

www.cellbiolabs.com

Cell Line

Catalog Number

info@cellbiolabs.com

123

CELL SIGNALING & PROTEIN BIOLOGY Reporter Molecules Recombinant Fluorescent Proteins Recombinant EGFP and RFP are provided at a concentration of 1 mg/mL and includes a 6xHis-tag at the C-terminus.

Recent Product Citation Sokolova, E. et al. (2013). Enhanced transcription rates in membrane-free protocells formed by coacervation of cell lysate. PNAS 110:11692-11697. (STA-201)

Product Name Recombinant EGFP

Recombinant RFP

Size

Catalog Number

100 µg

STA-201

5 x 100 µg

STA-201-5

100 µg

STA-202

5 x 100 µg

STA-202-5

Monoclonal Antibodies to Reporter Molecules Antibodies are provided at a concentration of 1 mg/mL. GFP antibody also recognizes EGFP, YFP, EYFP and CFP. RFP antibody recognizes Tag-RFP, Turbo-RFP, DeRed, mCherry and mOrange. Antibodies are suitable for Western blot, Immunostaining, ELISA and Dot blot. Recent Product Citation Maamary, J. et al. (2012). Attenuated influenza virus constructs with enhanced hemagglutinin protein expression. J. Virol. 86:5782-5790. (AKR-021) Product Name

Size

Catalog Number

Mouse Anti-GFP Monoclonal Antibody (clone GF28R)

100 µg

AKR-020

Mouse Anti-RFP Monoclonal Antibody (clone RF5R)

100 µg

AKR-021

Reporter Viral Vectors Premade viruses and viral constructs with reporter molecules make ideal controls for viral gene delivery experiments.

Reporter AAV Product Name

Reporter Adenoviruses Catalog Number

Product Name

Catalog Number

AAV1-GFP Control Virus

AAV-301

-Galactosidase

ADV-002

AAV2-Cre Control Virus

AAV-310

Firefly Luciferase

ADV-008

AAV2-GFP Control Virus

AAV-302

GFP

ADV-004

AAV2-Luc Control Virus

AAV-320

AAV3-GFP Control Virus

AAV-303

AAV5-GFP Control Virus

AAV-305

AAV6-GFP Control Virus

AAV-306

Reporter Retroviral Plasmids Target Name

Vector Backbone

Catalog Number

GFP

pBABE

RTV-002

GFP

pMCs

RTV-051

Catalog Number

GFP

pMX

RTV-050

GFP Lentivirus Control

LTV-300

GFP

pMYs

RTV-052

RFP Lentivirus Control

LTV-301

GFP-Puro

pMX

RTV-053

Reporter Lentiviruses Product Name

124

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Epitope Tags/Antibodies CELL SIGNALING & PROTEIN BIOLOGY His-Tag Protein ELISA Kit Our His-Tag Protein ELISA Kit allows you to detect and quantify His-tagged protein samples simply and reliably by comparing unknown samples with a recombinant standard. The kit is suitable for use with cell lysates and tissue homogenates.  Sensitive: Detect as little as 1 ng/mL protein or 50 pM of 6xHis-tag residues  Versatile: Use with proteins containing His-tag at either N- or C-terminus Product Name His-Tag Protein ELISA Kit

Recent Product Citation Dong, Y. et al. (2013). HMGB1 protein does not mediate the inflammatory response in spontaneous spinal cord regeneration: A hint for CNS regeneration. J. Biol. Chem. 288:18204-18218.

Detection

Size

Catalog Number

Colorimetric

96 Assays

AKR-130

Monoclonal Antibodies to Epitope Tags Antibodies are provided at a concentration of 1 mg/mL. GAPDH, ß-Actin and ß-Tubulin are also available as loading controls. All are suitable for Western blot, Immunostaining, ELISA, Immunoprecipitation, and Dot blot. Product Name

Size

Catalog Number

Mouse Anti-FLAG Tag Monoclonal Antibody (clone FG4R)

100 µg

AKR-004

Mouse Anti-GST Tag Monoclonal Antibody (clone GST.B6)

100 µg

AKR-005

Mouse Anti-HA Tag Monoclonal Antibody (clone HA.C5)

100 µg

AKR-006

Mouse Anti-His Tag Monoclonal Antibody (clone HIS.H8)

100 µg

AKR-003

Mouse Anti-Myc Tag Monoclonal Antibody (clone Myc.A7)

100 µg

AKR-007

Mouse Anti-V5 Tag Monoclonal Antibody (clone E10)

100 µg

AKR-008

Mouse Anti-GAPDH Monoclonal Antibody

100 µg

AKR-001

Mouse Anti-ß-Actin Monoclonal Antibody

100 µg

AKR-002

Mouse Anti-ß-Tubulln Monoclonal Antibody

100 µg

AKR-009

Rapid Antibody Purification Kit Our Rapid Antibody Purification Kit is designed for fast, single-step purification of high-quality IgG from ascites, serum, tissue culture media or hybridoma supernatants. IgG-containing samples are incubated with immobilized Protein A in the presence of a binding buffer. Non-IgG components are washed and IgG is subsequently eluted. The supplied Protein A column is suitable for 10 purification preps. The capacity per prep depends on the species of IgG; examples are listed in the column at right.

Species

mg of IgG per Prep

Bovine

15-20

Goat

6-12

Human

20-30

Mouse

6-12

Rabbit

15-20

Capacity per Prep for the Rapid Antibody Purification Kit.

Product Name Rapid Antibody Purification Kit

www.cellbiolabs.com

Size

Catalog Number

10 Preps

AKR-160

info@cellbiolabs.com

125

CELL SIGNALING & PROTEIN BIOLOGY Protein Phosphorylation PhosphoBLOCKER™ Western Blot Blocking Reagent Western blot blockers such as dry milk or serum are sufficient to block unreactive sites on the membrane. However, they are not designed to preserve phosphoprotein antigens during blotting. Recent Product Citations 1. Marques, J. et al. (2013). CRMP2 tethers kainate receptor activity to cytoskeleton dynamics during neuronal maturation. J. Neurosci. 33:18298-18310. 2. Ferreira, E. et al. (2013). Inflammatory cytokines induce a unique mineralizing phenotype in mesenchymal stem cells derived from human bone marrow. J. Biol. Chem. 288:2955029561. 3. Rosich, L. et al. (2012). Counteracting autophagy overcomes resistance to everolimus in mantle cell lymphoma. Clin. Cancer Res. 18:5278-5289. 4. Martinez, P. et al. (2012). 53BP1 deficiency combined with telomere dysfunction activates ATR-dependent DNA damage response. J. Cell Biol. 197:283-300.

 High Sensitivity: Enhances low level phosphoprotein signal without increasing background  Easy-to-use: Premixed dry blend

Dry Milk

PhosphoBLOCKER™ Reagent Superior Blocking with PhosphoBLOCKER™ Reagent. A549 cell lysate was blocked with dry milk or PhosphoBLOCKER before detection with anti-Phospho-p38 antibody.

Product Name PhosphoBLOCKER™ Western Blot Blocking Reagent

Size

Catalog Number

AKR-103

AKR-104

Phospho Antibody Stripping Solution This solution removes anti-phosphoantibodies selectively from blots without significantly affecting the immobilized proteins, allowing re-probing of the blot with the same or a different antibody. Stripping of antibodies is done at room temperature, so no heating of blots is required. Product Name Phospho Antibody Stripping Solution

Multiple Blotting and Stripping of 4G10 Phosphotyrosine Antibody.

Size

Catalog Number

10 mL

AKR-102

Phosphoprotein Purification Kit Our Phosphoprotein Purification Kit allows you to enrich your phosphoprotein samples quickly and easily. Phosphorylated proteins are affinity purified from lysates with a single-step purification / enrichment matrix. The entire procedure takes about 4 hours. Each prep can process 2.5 mg of total lysate protein, or approximately one confluent 10 cm dish.

Product Name Phosphoprotein Purification Kit

126

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Enrichment of p-ERK. HeLa cell lysate was incubated with the Phosphoprotein Enrichment Matrix from the Phosphoprotein Purification Kit.

Size

Catalog Number

2 preps

AKR-105

5 preps

AKR-106

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Protein Isolation CELL SIGNALING & PROTEIN BIOLOGY Rapid GST Inclusion Body Solubilization and Renaturation Kit The Rapid GST Inclusion Body Solubilization and Renaturation Kit is designed to retrieve expressed GST fusion proteins in soluble form after lysis and extraction. Each kit contains sufficient reagents to solubilize and renature up to 5-10 liters of bacterial culture.

Pre Beads Post Beads

GS Beads

1 2 3 4 5 6 7 8 9 10 11 12 13 14

 Fast Results: No lengthy dialysis or dilution steps  Easy-to-Use: No pH variation or redox pair  Optimized: Designed specifically to solubilize and renature GST inclusion bodies Recent Product Citations 1. Keller, D. et al. (2014). Mechanisms of HsSAS-6 assembly promoting centriole formation in human cells. J. Cell Biol. 204:697712. 2. Lalani, S. et al. (2013). MCTP2 is a dosage-sensitive gene required for cardiac outflow tract development. Hum. Mol. Genet. 10.1093/hmg/ddt283.

Solubilization and Renaturation of GST-RTK Fusion Protein. Lane 1: MW STD; Lane 2: Whole E.Coli lysate; Lane 3, 7, 11: No detergent; Lane 4, 8, 12: 32-fold dilution; Lane 5, 9, 13: 8-fold dilution; Lane 6, 10, 14: 2-fold dilution.

Product Name

Size

Catalog Number

Rapid GST Inclusion Body Solubilization and Renaturation Kit

1 Kit

AKR-110

Nuclear/Cytosolic Fractionation Kit The Nuclear/Cytosolic Fractionation Kit provides a simple and fast tool to isolate nuclear extract from the cytoplasmic fraction of mammalian cells. The optimized protocol provides high protein recovery and low cross-contamination in less than 2 hours. Extracted fractions are functional and suitable for downstream assays such as DNA footprinting, pre-mRNA splicing, gel shift assays, reporter assays, enzyme activity assays, and Western blot.

Product Name Nuclear/Cytosolic Fractionation Kit

HEK293 Cell Fractionation. Whole cell (W), cytosolic (C), and nuclear (N) fractions were immunoblotted with Anti-Tubulin (cytosol specific) or Anti-Lamin A/C (nuclear specific).

Size

Catalog Number

20 preps

AKR-171

100 preps

AKR-172

Bacterial Protein Extraction Reagents Our Bacterial Protein Extraction Reagents contain a gentle, non-ionic detergent formulation that quickly extracts functional recombinant proteins from E. coli without mechanical disruption. The reagents are compatible with 6xHis and GST affinity purification systems and are suitable for small-scale or large-scale protein extraction. Product Name

Size

Catalog Number

5X Bacterial Protein Extraction Reagent (Phosphate)

50 mL

AKR-181

5X Bacterial Protein Extraction Reagent (Tris)

50 mL

AKR-180

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127

CELL SIGNALING & PROTEIN BIOLOGY Protein Quantitation High Sensitivity Protein Quantitation Assay Kit Our High Sensitivity Protein Quantitation Assay Kit provides a robust method to quantify protein concentrations in the µg/mL range. The fluorometric format creates a significant sensitivity advantage over standard colorimetric methods such as Bradford or BCA assays.

 Highly Sensitive: Quantify protein concentrations as low as 5 µg/mL  Fast: Obtain results in 5 to 15 minutes  Easy-to-use: Read on a 96-well fluorescence plate reader

250

150

RFU

200

100

50 0

100

200

300

400

500

BSA Standard (ug/mL)

Standard Curve Generated with the High Sensitivity Protein Quantitation Assay Kit. Product Name High Sensitivity Protein Quantitation Assay Kit

Size

Catalog Number

100 Assays

AKR-185

RIPA Buffer RIPA buffer is a popular reagent for lysis of both adherent and suspension cells in culture, as well as making tissue homogenates. RIPA buffer extracts cytoplasmic, membrane, and nuclear proteins for a variety of downstream protein assays and immunoassays. Our RIPA Buffer is provided as a 5X concentrate and is available with or without a Protease Inhibitor Cocktail.

128

Product Name

Size

Catalog Number

5X RIPA Buffer

20 mL

AKR-191

5X RIPA Buffer, with Protease Inhibitor Cocktail

20 mL

AKR-190

Phone 1-858-271-6500

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Fax 1-858-271-6514

METABOLISM RESEARCH

Lipoprotein Metabolism

Lipoprotein Metabolism Research Lipoproteins are important assemblies of proteins and lipids that have implications in cardiovascular and related diseases. Our kits and reagents can help streamline the study of various members of lipoprotein metabolism pathways:  Cholesterol / Lipoproteins  Lipoprotein Lipase  Apolipoproteins  Triglycerides  Oxidized LDL  Free Fatty Acids  Lipoprotein Receptors  Phospholipids  Cholesteryl Ester Pathway  Serum Proteins

Total Cholesterol Assay Kits Cholesterol exists within lipoproteins in two forms: a free alcohol and a fatty cholesteryl ester, which is the predominant form of cholesterol transport and storage.

 Sensitive: Detect as little as 100 nM  Fast: Simple 30 minute protocol  Easy-to-use: Detect in a 96-well plate reader; colorimetric and fluorometric formats available

Our Total Cholesterol Assay Kits provide a simple plate-based format that measures the amount of cholesterol in serum, plasma, cell lysates or tissue homogenates. In the presence of cholesterol esterase, the assay will measure total cholesterol in both forms. In the absence of the esterase, the assay will measure only free cholesterol.

Recent Product Citations 1. Marino, A. et al. (2014). ITCH deficiency protects from dietinduced obesity. Diabetes 63:550-561. (STA-384) 2. Mathews, E. et al. (2014). Mutation of 3-hydroxy-3methylglutaryl CoA synthase I reveals requirements for isoprenoid and cholesterol synthesis in oligodendrocyte migration arrest, axon wrapping, and myelin gene expression. J. Neurosci. 34:3402-3412. (STA-384)

Quantitation is performed in a 96-well plate reader with your choice of colorimetric or fluorescence-based detection. 5000

Plasma

4500

Serum

4000 3500 3000 s U2500 F R 2000 1500 1000 500 0 1 to 10

1 to 100 1 to 1000 Serum and Plasma Dilutions

1 to 10000

Cholesterol Values from Normal Human Serum and Plasma Samples. Relative Fluorescence Unit (RFU) values are then compared against a Cholesterol Standard Curve (not shown).

Product Name Total Cholesterol Assay Kit

130

Phone 1-858-271-6500

Assay Principle for the Total Cholesterol Assay Kit (Fluorometric). Detection

Size

Catalog Number

Colorimetric

192 Assays

STA-384

Fluorometric

192 Assays

STA-390

USA Toll-Free 1-888-CBL-0505

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METABOLISM RESEARCH

Lipoprotein Metabolism HDL-Cholesterol Assay Kit Our HDL-Cholesterol Assay Kit provides a simple plate-based format similar to our Total Cholesterol Assays (previous page). This kit measures the amount of cholesterol in the HDL fraction isolated from serum, plasma, cell lysates or tissue homogenates.

 Sensitive: Detect as little as 1 µM  Fast: Simple 45 minute assay protocol  Easy-to-use: Detect in a 96-well plate fluorescence-based plate reader

Product Name HDL-Cholesterol Assay Kit

Detection

Size

Catalog Number

Fluorometric

96 Assays

STA-394

HDL and LDL/VLDL Cholesterol Assay Kit

In the presence of cholesterol esterase, the assay will measure total cholesterol in both free cholesterol and cholesteryl ester forms. In the absence of the esterase, the assay will measure only free cholesterol. Quantitation of cholesteryl ester alone may be calculated by subtracting free cholesterol from total cholesterol levels. Product Name HDL and LDL/VLDL Cholesterol Assay Kit

8000

7037

6000 RFU

Our HDL and LDL/VLDL Cholesterol Assay Kit is similar in principle to our Total Cholesterol Assay Kit, but allows you to quantify HDL and LDL/VLDL separately in serum samples. After separating samples into HDL and LDL/VLDL fractions, the fluorometric assay is run according to the Assay Principle for the Total Cholesterol Assay Kit (previous page).

Total Cholesterol (HDL + LDL/VLDL)

5011

LDL/VLDL

HDL

4000

1769

2000 0 1

Quantitation of Total Cholesterol, LDL/VLDL, and HDL.

Detection

Size

Catalog Number

Fluorometric

192 Assays

STA-391

Human Lipoproteins Our human lipoproteins are isolated from human plasma following ultracentrifugation.

Product Name

Size

Catalog Number

Human High Density Lipoprotein (HDL)

100 µg

STA-243

Human High Density Lipoprotein-2 (HDL-2)

100 µg

STA-244

Human High Density Lipoprotein-3 (HDL-3)

100 µg

STA-245

Human Low Density Lipoprotein (LDL)

100 µg

STA-241

Human Low Density Lipoprotein (LDL), Copper (Cu++) Oxidized

100 µg

STA-214

Human Low Density Lipoprotein (LDL), Malondialdehyde Modified

100 µg

STA-212

Human Low Density Lipoprotein (LDL), Nitrated

100 µg

STA-213

Human Very Low Density Lipoprotein (VLDL)

100 µg

STA-242

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131

METABOLISM RESEARCH

Lipoprotein Metabolism

Human Apolipoprotein ELISA Kits Apolipoproteins comprise the protein component of lipoprotein assemblies. Apolipoproteins fall into two classes:  Non-exchangeable Apo B associates with LDL  Exchangeable Apo A, C and E associate with HDL Our Human Apo ELISA Kits provide a convenient and sensitive method for quantifying specific apolipoproteins in serum, plasma, or other biological fluids.

Apo (a)

Apo AI

Apo Apo Apo Apo Apo AII B CI CII CIII

Apo E

1 ng/mL

50 pg/mL

0.3 ng/mL

200 pg/mL

1 200 1 ng/mL pg/mL ng/mL

50 pg/mL

Detection Limits of Cell Biolabs’ Human Apo ELISA Kits.

Product Name

Human ApoB ELISA Standard Curve.

Detection

Size

Catalog Number

Human Apo(a) ELISA Kit

Colorimetric

96 Assays

STA-359

Human ApoAI ELISA Kit

Colorimetric

96 Assays

STA-362

Human ApoAII ELISA Kit

Colorimetric

96 Assays

STA-363

Human ApoB ELISA Kit

Colorimetric

96 Assays

STA-368

Human ApoCI ELISA Kit

Colorimetric

96 Assays

STA-364

Human ApoCII ELISA Kit

Colorimetric

96 Assays

STA-365

Human ApoCIII ELISA Kit

Colorimetric

96 Assays

STA-366

Human ApoE ELISA Kit

Colorimetric

96 Assays

STA-367

Human Apolipoproteins Following ultracentrifugation, lipoproteins are isolated from human plasma. Water-soluble apolipoproteins are then purified from delipidated lipoprotein. Product Name

132

Size

Catalog Number

Human Albumin, Malondialdehyde Modified

100 µg

STA-210

Human Apolipoprotein AI

100 µg

STA-232

Human Apolipoprotein AII

100 µg

STA-233

Human Apolipoprotein B-100

100 µg

STA-234

Human Apolipoprotein B-100, Malondialdehyde Modified

100 µg

STA-211

Human Apolipoprotein CI

100 µg

STA-235

Human Apolipoprotein CIII

100 µg

STA-237

Phone 1-858-271-6500

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Lipoprotein Metabolism

METABOLISM RESEARCH

Human ApoAI and ApoB Duplex ELISA Kit As the primary protein components of HDL and LDL respectively, ApoAI and ApoB are arguably the most significant apolipoproteins in lipid metabolism research. Our Human ApoAI and ApoB Duplex ELISA Kit provides a convenient tool to quantify both proteins in a single serum or plasma sample. Unlike other multiplex assays, our ApoAI and ApoB Duplex ELISA does not require a luminometer for detection. Simply quantify both proteins using a standard colorimetric ELISA plate reader. The ELISA plate is coated with Anti-ApoAI and AntiApoB antibodies, which respectively capture HDL and LDL from the sample. Biotinylated Anti-ApoAI and Anti-ApoB detection antibodies are added, followed by enzyme conjugates. Alkaline phosphatase substrate is added allowing quantitation of ApoAI. After washing, HRP is added to allow quantiation of ApoB.

 Efficient: Quantify two apolipoproteins from the same sample in just a few hours  Sensitive: Detect as little as 0.1 ng/mL of ApoAI and 1 ng/mL of ApoB from serum or plasma  Quantitative: Compare results to known ApoAI and ApoB standards Human ApoAI and ApoB Duplex ELISA Kit Assay Principle. Product Name

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-361

Detection

Size

Catalog Number

Sheep Anti-Human Apolipoprotein (a) Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-131

Goat Anti-Human Apolipoprotein AI Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-132

Rabbit Anti-Human Apolipoprotein AII Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-133

Goat Anti-Human Apolipoprotein B-100/48 Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-134

Rabbit Anti-Human Apolipoprotein CI Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-135

Rabbit Anti-Human Apolipoprotein CII Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-136

Rabbit Anti-Human Apolipoprotein CIII Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-137

Goat Anti-Human Apolipoprotein E Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-138

Human ApoAI and ApoB Duplex ELISA Kit

Antibodies to Apolipoproteins Antibodies are affinity purified. Product Name

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133

METABOLISM RESEARCH

Lipoprotein Metabolism

Quantitation of MDA-LDL in Serum and Plasma Samples. Serum and plasma samples were treated with LDL Precipitation Solution. Precipitated LDL pellets were resuspended in 1.6 mL of PBS before further dilution 1:160 in Assay Diluent according to the Assay Protocol.

OxiSelect™ Human Oxidized LDL ELISA Assay Principle.

134

Detection

Size

Catalog Number

OxiSelect™ Human Oxidized LDL ELISA Kit (CML-LDL Quantitation)

Colorimetric

96 Assays

STA-388

OxiSelect™ Human Oxidized LDL ELISA Kit (HNE-LDL Quantitation)

Colorimetric

96 Assays

STA-389

OxiSelect™ Human Oxidized LDL ELISA Kit (MDA-LDL Quantitation)

Colorimetric

96 Assays

STA-369

OxiSelect™ Human Oxidized LDL ELISA Kit (OxPL-LDL Quantitation)

Colorimetric

96 Assays

STA-358

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Lipoprotein Metabolism

METABOLISM RESEARCH

Human LDL Receptor ELISA Kit Cholesterol can be toxic when accumulated in excess in cell membranes. The low-density lipoprotein receptor (LDLR) is the primary means of removing cholesterol from the circulation. LDLR is a transmembrane protein that transports cholesterol-carrying lipoprotein particles (primarily LDL) into cells. Receptor-ligand complexes enter the cell by endocytosis; bound lipoproteins are subsequently released in the low-pH setting of the endosome, while the receptors return to the cell surface. Our Human LDLR ELISA Kit provides a simple, convenient method for the detection and quantitation of LDL receptor in a variety of human sample types.  Sensitive: Detect as little as 50 pg/mL of human LDLR  Versatile: Assay is compatible with plasma, serum, cell lysates and tissue homogenates  Quantitative: Compare results to a known human LDLR standard Product Name Human LDLR ELISA Kit

Standard Curve Generated with the Human LDLR ELISA Kit.

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-386

Human LOX-1 ELISA Kit Monocytes and macrophages can form atherosclerotic lesions when they take in oxidized LDL (OxLDL). Uptake is done via the lectin-like oxidized LDL receptor-1 (LOX-1), which is expressed in vascular endothelium as well as in vascular smooth muscle cells, differentiated macrophages and platelets. LOX-1 can be cleaved and released as a soluble form (sLOX-1), which can serve as a prognostic biomarker in serum for early acute coronary syndromes, stroke and coronary heart disease.

Recent Product Citation Wu, J. et al. (2013). Clinical nephrology—IgA nephropathy, lupus nephritis, vasculitis. Nephrol. Dial. Transplant. 28:i175-i184.

Our Human LOX-1 ELISA Kit provides a simple, convenient method for the detection and quantitation of LOX-1 receptor in a variety of sample types.  Sensitive: Detect as little as 40 pg/mL of human LOX-1  Versatile: Assay is compatible with plasma, serum, cell lysates, tissue homogenates, or cell culture supernatants  Quantitative: Compare results to a known human LOX-1 standard Product Name Human LOX-1 ELISA Kit

Standard Curve Generated with the Human LOX-1 ELISA Kit. Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-387

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135

METABOLISM RESEARCH

Lipoprotein Metabolism

Human PCSK9 ELISA Kit Proprotein convertase subtilisin kexin 9 (PCSK9) is a member of the proteinase K subfamiliy of subtilisinrelated serine endoproteases. PCKS9 mediates LDL receptor (LDLR) degradation by binding to the EGF domain of the LDLR. This binding prevents LDLR from being sorted to the endosomes for recycling back to the cell surface. Instead, the PCSK9/LDLR complex is distributed to the lysosomes for degradation. Our Human PCKS9 ELISA Kit provides a simple, convenient method for the detection and quantitation of PCSK9 in a variety of human sample types.  Sensitive: Detect as little as 150 pg/mL of human PCSK9  Versatile: Assay is compatible with plasma, serum, and cell and tissue lysates  Quantitative: Compare results to a known human PCSK9 standard Product Name Human PCSK9 ELISA Kit

Standard Curve Generated with the Human PCSK9 ELISA Kit.

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-385

Human Cholesteryl Ester Transfer Protein (CETP) ELISA Kit Cholesterol exists within lipoproteins in two forms: a free alcohol and a fatty cholesteryl ester. The cholesteryl ester is the predominant form of cholesterol transport and storage. Cholesteryl ester transfer protein (CETP) promotes the transfer of both cholesteryl esters and triglycerides between various types of lipoprotein particles: HDL, LDL, VLDL, and chylomicrons. Our Human CETP ELISA Kit provides a simple, convenient method for the detection and quantitation in a variety of human sample types.  Sensitive: Detect as little as 60 ng/mL of human CETP  Versatile: Assay is compatible with plasma, serum, and other biological fluids  Quantitative: Compare results to a known human CETP standard Product Name Human Cholesteryl Ester Transfer Protein (CETP) ELISA Kit

136

Phone 1-858-271-6500

CETP Promotes Bidirectional Transfer of Cholesteryl Esters (CE) and Triglycerides (TG) Between Lipoproteins. Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-614

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Lipoprotein Metabolism

METABOLISM RESEARCH

Lecithin Cholesterol Acyltransferase (LCAT) Activity Assay Kit Lecithin cholesterol acyltransferase (LCAT) is an enzyme that is associated with lipoproteins and plays a key role in promoting the transfer of excess cellassociated cholesterol from peripheral tissues to the liver to be excreted. LCAT catalyzes the transfer of an sn-2 acyl group from phosphatidylcholine to cholesterol, forming a cholesteryl ester. LCAT is bound to various lipoproteins in the blood, including HDL and LDL. Our LCAT Activity Assay Kit provides a simple, convenient method for measuring the phospholipase activity of LCAT in a variety of sample types including plasma, serum, cell lysates and tissue homogenates. Quantitation of LCAT activity is performed in a 96well fluorescence-based plate reader. This assay may also be used to quantify other calcium independent phospholipase activities such as lipoprotein phospholipase A2 (LP-PLA2). Product Name Lecithin Cholesterol Acyltransferase (LCAT) Activity Assay Kit

Assay Principle for the LCAT Activity Assay Kit. The close proximity of fluorescence labels on a dual-labeled fluorogenic probe keeps the fluorescence quenched. Upon cleavage of the probe by LCAT, fluorescence of the monomers can be measured at an excitation of 342 nm and emission of 400 nm. Detection

Size

Catalog Number

Fluorometric

100 Assays

STA-615

Lecithin Cholesterol Acyltransferase (LCAT) ELISA Kit Lecithin cholesterol acyltransferase (LCAT) is an enzyme that is associated with lipoproteins and plays a key role in promoting the transfer of excess cellassociated cholesterol from peripheral tissues to the liver to be excreted. LCAT catalyzes the transfer of an sn-2 acyl group from phosphatidylcholine to cholesterol, forming a cholesteryl ester. LCAT is bound to various lipoproteins in the blood, including HDL and LDL. Our LCAT ELISA Assay Kit provides a simple, convenient method for quantifying LCAT levels in a variety of sample types including plasma, serum, and cell and tissue lysates. Quantitation of LCAT activity is performed in a standard 96-well plate reader.  Sensitive: Detect as little as 30 ng/mL of LCAT  Versatile: Suitable for human, mouse, rat or rabbit samples Product Name Lecithin Cholesterol Acyltransferase (LCAT) ELISA Kit

Standard Curve Generated with the Lecithin Cholesterol Acyltransferase (LCAT) ELISA Kit.

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-616

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137

METABOLISM RESEARCH

Lipoprotein Metabolism

Lipoprotein Lipase (LPL) Activity Assay Kit Lipoprotein lipase (LPL) is the key plasma lipase responsible for the hydrolysis of the triglyceride core found in very low density lipoprotein (VLDL) particles formed in the liver. Our Lipoprotein Lipase (LPL) Activity Assay Kit provides a simple, convenient method to measure LPL activity in a variety of sample types. This kit uses a fluorogenic triglyceride analog as a lipase substrate. The quenched substrate is cleaved at the sn-1 position by LPL producing a fluorescent product that can be detected in a 96-well fluorescence plate reader. This assay will also measure the activity of endothelial and hepatic lipases. It cannot distinguish between these lipases and LPL. Recent Product Citations 1. Dib, L. et al. (2014). LXRalpha fuels fatty acid-stimulated oxygen consumption in white adipocytes. J. Lipid Res. 55:247-257. 2. Navab, M. et al. (2013). Transgenic 6F tomatoes act on the small intestine to prevent systemic inflammation and dyslipidemia caused by Western diet and intestinally derived lysophosphatidic acid. J. Lipid Res. 54:3403-3418.

Product Name Lipoprotein Lipase (LPL) Activity Assay Kit

Assay Principle for the LPL Activity Assay Kit. The fluorogenic substrate is initially quenched and non-fluorescent. Upon cleavage of the probe by incubation with lipoprotein lipase (LPL), fluorescence can be measured at an excitation of 342 nm and emission of 400 nm. Detection

Size

Catalog Number

Fluorometric

100 Assays

STA-610

Lipoprotein Lipase (LPL) ELISA Kit Lipoprotein lipase (LPL) is the key plasma lipase responsible for the hydrolysis of the triglyceride core found in very low density lipoprotein (VLDL) particles formed in the liver. A mutation in the gene coding for LPL can lead to deficiencies in the enzyme, resulting in a diminished ability to breakdown fatty acids. Such LPL deficiency is known as chylomicronemia or Type I hyperlipoproteinemia. Our Lipoprotein Lipase (LPL) ELISA Kit provides a simple, convenient method to measure LPL levels in plasma, serum or other biological fluids from a variety of species (see below). LPL amounts are quantified against the provided LPL Standard in a colorimetric 96-well microplate reader.  Sensitive: Detect as little as 20 ng/mL of LPL  Versatile: Suitable for human, rat, bovine, guinea pig, or chicken samples (but not mouse) Product Name Lipoprotein Lipase (LPL) ELISA Kit

138

Phone 1-858-271-6500

Standard Curve Generated with the Lipoprotein Lipase (LPL) ELISA Kit.

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-611

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Lipoprotein Metabolism

METABOLISM RESEARCH

Serum Triglyceride Quantitation Kits Triglycerides serve as an energy source and play a key role in lipid metabolism. Lipases secreted into the intestines hydrolyze the triglyceride ester bond, producing glycerol and free fatty acids. Hepatic lipases also break down triglycerides in the liver to assemble very low density lipoprotein (VLDL) particles.

 Sensitive: Detect as little as 10 µM (1 mg/dL) with the colorimetric format and 2 µM (0.2 mg/dL) with the fluorometric format  Versatile: Suitable for serum, plasma, and cell and tissue lysates

Our Serum Triglyceride Quantitation Kits use a coupled enzymatic reaction system to measure triglyceride concentrations. First, a lipase hydrolyzes the ester bond, yielding free glycerol. The glycerol is then phosphorylated and oxidized, producing hydrogen peroxide, which reacts with the probe provided with each kit. Kits are available with either colorimetric or fluorescence-based detection, both of which are performed in a 96-well microtiter plate. Recent Product Citations 1. Chellan, B. et al. (2014). IL-22 is induced by S100/calgranulin and impairs cholesterol efflux in macrophages by downregulating ABCG1. J. Lipid Res. 55:443-454. (STA-396) 2. Marino, A. et al. (2014). ITCH deficiency protects from dietinduced obesity. Diabetes 63:550-561. (STA-396)

Standard Curve Generated with the Serum Triglyceride Quantitation Kit (Colorimetric).

Want to measure free glycerol content? See our Free Glycerol Assay Kits on page 146. Product Name Serum Triglyceride Quantification Kit

Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-396

Fluorometric

100 Assays

STA-397

Free Fatty Acid (FFA) Assay Kits Free fatty acids (FFA) are released upon hydrolysis of triglycerides. FFAs then bind plasma albumin for circulation in the body, serving as a readily absorbed energy source for muscle, brain, and other organ tissues. Our Free Fatty Acid Assay Kits use a coupled enzymatic reaction system to measure free fatty acid concentrations in serum or plasma. Acyl CoA Synthetase catalyzes FFA acylation of CoA. The Acyl-CoA is then oxidized by Acyl CoA Oxidase, producing hydrogen peroxide, which reacts with the kit’s probe. Kits are available with either colorimetric or fluorescencebased detection, both of which are performed in a 96well microtiter plate. Product Name Free Fatty Acid Assay Kit

Standard Curve Generated with the Free Fatty Acid Assay Kit (Fluorometric).

Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-618

Fluorometric

100 Assays

STA-619

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139

METABOLISM RESEARCH

Lipoprotein Metabolism

Phosphatidylcholine Assay Kit Phosphatidylcholine is the foremost phospholipid in eukaryotic cell membranes and comprises about 70% of the total phospholipids in plasma lipoproteins. Our Phosphatidylcholine Assay Kit is a simple fluorometric assay that measures phosphatidylcholine in plasma, serum, cell suspensions or tissue homogenates. Phospholipase D enzyme hydrolyzes phosphatidylcholine into phosphatidic acid and choline. The choline is then oxidized by choline oxidase to produce hydrogen peroxide, which is detected by a fluorogenic probe in the presence of horseradish peroxidase (HRP).

Product Name Phosphatidylcholine Assay Kit

Detection

Size

Catalog Number

Fluorometric

96 Assays

STA-600

Sphingomyelin Assay Kit 3500 3000 2500

RFUs

Our Sphingomyelin Assay Kit is a simple fluorometric assay that measures sphingomyelin levels in plasma, serum, cell suspensions or tissue homogenates. Sphingomyelinase hydrolyzes sphingomyelin into ceramide and phosphocholine, which in turn is broken down into choline. Choline is enzymatically oxidized to produce hydrogen peroxide, which is detected with a fluorogenic probe in the presence of horseradish peroxidase (HRP).

2000 1500 1000 500

Recent Product Citation Winklger, E.A. et al. (2014). Blood-spinal cord barrier disruption contributes to early motor-neuron degeneration in ALS-model mice. PNAS 111:E1035-E1042.

0 0

10 15 SphingomyelinÂ (mg/dL)

Standard Curve Generated with the Sphingomyelin Assay Kit. Product Name Sphingomyelin Assay Kit

Detection

Size

Catalog Number

Fluorometric

96 Assays

STA-601

Acetylcholine Assay Kits Our Acetylcholine Assay Kits provide a simple, convenient method to quantify acetylcholine in plasma, serum, cell suspensions, or tissue homogenates. Kits are available with either colorimetric or fluorometric detection, both of which are performed in a 96-well microplate reader. Product Name Acetylcholine Assay Kit

140

Phone 1-858-271-6500

Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-603

Fluorometric

100 Assays

STA-602

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Lipoprotein Metabolism

METABOLISM RESEARCH

Human C-Reactive Protein ELISA Kit C-Reactive Protein (CRP) is a serum protein that binds with high affinity to phosphocholine residues as well as other autologous and extrinsic ligands. CRP is a well-established marker of inflammation and tissue damage, and it has been associated with cardiovascular disease, atherosclerosis, and other diseases. Our Human C-Reactive Protein (CRP) ELISA Kit provides a simple, convenient method to measure CRP levels in human plasma, serum, or other biological fluids. CRP amounts are quantified against the provided CRP Standard in a colorimetric 96-well microplate reader.

 Sensitive: Detect as little as 1 ng/mL of CRP  Versatile: Suitable for plasma, serum, or other biological fluids Product Name Human C-Reactive Protein (CRP) ELISA Kit

Standard Curve Generated with the Human C-Reactive Protein (CRP) ELISA Kit. Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-392

Human Plasminogen ELISA Kit Plasminogen is a plasma glycoprotein that plays a role in macrophage recruitment, arterial stenosis, atherosclerosis, aneurysm formation, wound healing, and neovascularization. Plasminogen exists as an inactive proenzyme, but when converted to the active enzyme plasmin it serves to digest fibrin. This activation is catalyzed by tissue-type plasminogen activator (tPA) or urokinase-type plasminogen activator (uPA). Our Human Plasminogen ELISA Kit provides a simple, convenient method to measure plasminogen levels in human plasma, serum, or other biological fluids. Plasminogen amounts are quantified against the provided Plasminogen Standard in a colorimetric 96well microplate reader.  Sensitive: Detect as little as 150 pg/mL of plasminogen  Versatile: Suitable for plasma, serum, or other biological fluids

Product Name Human Plasminogen ELISA Kit

Standard Curve Generated with the Human Plasminogen ELISA Kit.

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-393

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141

METABOLISM RESEARCH

Lipoprotein Metabolism

Human Albumin ELISA Kit Human serum albumin (HSA) is the most abundant protein in human plasma, constituting about half the protein in blood serum. It is typically found in concentrations around 50 mg/mL. Produced in the liver in a preproalbumin state, albumin transports hormones, fatty acids, and other compounds through the circulation. It also maintains pH and osmotic pressure.

 Sensitive: Detect as little as 100 pg/mL of human serum albumin  Versatile: Suitable for plasma, serum, or other biological fluids

Our Human Albumin ELISA Kit provides a simple, convenient method to measure HSA levels in human plasma, serum, urine, or other biological fluids. Human albumin amounts are quantified against the provided HSA Standard in a colorimetric 96-well microplate reader.

Standard Curve Generated with the Human Albumin ELISA Kit. Product Name Human Albumin ELISA Kit

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-383

Human Serum Proteins Our human albumin and oxidized albumin were isolated and purified by HPLC. C-reactive protein was isolated from human pleural fluid. Plasminogen was isolated and purified from human plasma following an ultracentrifugation procedure. Product Name

Size

Catalog Number

Human Albumin

100 µg

STA-230

Human Albumin, Malondialdehyde Modified

100 µg

STA-210

Human C-Reactive Protein

100 µg

STA-240

Human Plasminogen

100 µg

STA-239

Detection

Size

Catalog Number

Goat Anti-Human Albumin Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-130

Rabbit Anti-Human C-Reactive Protein Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-140

Goat Anti-Human Plasminogen Polyclonal Antibody

Immunoblot/ELISA

100 µg

STA-139

Antibodies to Human Serum Proteins Antibodies are affinity purified. Product Name

142

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Lipoprotein Metabolism

METABOLISM RESEARCH

Lipid Extraction Kit, Chloroform Free Traditional methods of extracting lipids from tissues or cells, such as the well-published Folch method, have used chloroform as the extraction solvent. This method has a couple of disadvantages. First, the organic phase ends up below the aqueous phase, creating problematic removal through the upper phase that risks contamination. Second, chloroform has been classified in many places are a probable human carcinogen. Our Lipid Extraction Kit overcomes both disadvantages of the Folch method. The kit provides a chloroform-free extraction method, and the use of proprietary organic solvents places the organic phase above the aqueous phase, allowing easy removal without disturbing the aqueous layer. Each kit provides sufficient reagents for 50 extractions from 100 ÂľL sample sizes, but reagents may be scaled up for larger samples.

Total Cholesterol Assay Performed on Extracted Lipids. Lipids extracted from fetal bovine serum (FBS) and HEK293 cells were prepared using the traditional Folch method (blue) and the Lipid Extraction Kit (red). Samples were tested for the presence of cholesterol in the Total Cholesterol Assay Kit (Cat. #STA-390).

Product Name Lipid Extraction Kit (Chloroform Free)

Size

Catalog Number

50 Preps

STA-612

Lipid Quantification Kit Our Lipid Quantification Kit provides a convenient plate-based method to measure the total unsaturated lipid content found in various samples.* The assay uses a sulfo-phospho-vanillin method in which samples are acidified and heated to solubilize and prime the lipids. The lipids then react with vanillin in acidic conditions to form a colorimetric product detectable in a standard 96-well microplate reader. This kit is compatible with plasma and serum samples, or with crude or purified lipids extracted from cells. Each kit provides sufficient reagents for 100 assays including standards and unknown samples.

*Saturated lipids are not detected with this assay. Product Name Lipid Quantification Kit

Standard Curve Generated with the Total Lipid Quantification Kit. Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-613

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143

METABOLISM RESEARCH

Renal Function Assays

Renal Function Assays Our Renal Function Assays provide a simple, sensitive method to test for various markers related to kidney function:  

Uric Acid / Uricase Urea

Creatinine Protein Carbamylation

 

Uric Acid/Uricase Assay Kit Uric acid is the final oxidation end product of purine nucleotide metabolism. Uric acid is a potent antioxidant that is released during hypoxic conditions and is usually excreted in the urine via glomerular filtration. In the urine, the enzyme uricase metabolizes uric acid to allantoin which is excreted from the body. Our Uric Acid / Uricase Assay Kit is a simple 96-well microplate-based assay for measuring concentrations of either uric acid or uricase in serum, plasma or urine samples. Detection is performed in a fluorescence-based plate reader.  Sensitive: Detect as little as 0.5 µM of uric acid or 1 mU/mL of uricase  Quantitative: Kit includes both uric acid and uricase standards Assay Principle for the Uric Acid / Uricase Assay Kit. Product Name Uric Acid/Uricase Assay Kit

Detection

Size

Catalog Number

Fluorometric

400 Assays

STA-375

Urea Assay Kit

Our Urea Assay Kit is a simple 96-well microplatebased assay for measuring urea concentrations in serum, plasma, lysates, or urine samples. Detection is performed in standard colorimetric plate reader.  Sensitive: Detect as little as 1 mg/dL of urea  Quantitative: Measure unknown samples against a known urea standard curve

144

0.9 0.8 0.7 OD 630 nm

Urea is the end product of protein nitrogen metabolism and is the primary vehicle for removing toxic ammonia from the body. Urea quantitation is one of the most widely applied tests for kidney function evaluation.

0.6 0.5 0.4

Plasma

0.3

Serum

0.2 0.1 0 1 to 2

1 to 4 1 to 8 Plasma and Serum Dilutions

Human Plasma and Serum Samples Tested with the Urea Assay Kit.

Product Name

Detection

Size

Catalog Number

Urea Assay Kit

Fluorometric

192 Assays

STA-382

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

Renal Function Assays

METABOLISM RESEARCH

Urinary Creatinine Assay Kit Creatinine is a metabolite formed from creatine and phosphocreatine (p-creatine). Subsequently creatinine enters the blood and is then excreted by the kidneys via glomerular filtration. Intra-individual variation of creatinine levels is <15% daily, making it a useful marker for normalizing levels of other molecules found in the urine. Our Urinary Creatinine Assay Kit is based on the Jaffe reaction between creatinine and alkaline picrate, which produces an orange-red color complex that can be easily read by a standard microplate reader. A creatinine standard is provided to allow quantitative measurements of creatinine levels in urine samples. The assay is simple and takes less than one hour to perform. Product Name Urinary Creatinine Assay Kit

Creatinine Levels in Urine Samples in the Presence and Absence of Glucose.

Detection

Size

Catalog Number

Colorimetric

192 Assays

STA-378

Protein Carbamylation ELISA Kit and Antibodies Carbamylation is a post-translational modification which occurs throughout the lifespan of proteins in vivo. Carbamylation results from the binding of isocyanic acid, which spontaneously arises from high concentrations of urea, to lysine residues of proteins as carbamyl-lysine (CBL). Our Protein Carbamylation Sandwich ELISA Kit is a convenient microplate-based method for the evaluation of protein carbamylation in a variety of sample types. In addition, polyclonal antibodies are available for use in Western blot and ELISA applications.

Formation of Carbamyl-Lysine (CBL) During Carbamylation of Proteins.

Product Name

Standard Curve Generated with the OxiSelect™ Protein Carbamylation Sandwich ELISA Kit.

Detection

Size

Catalog Number

Colorimetric

96 Assays

STA-877

Goat Anti-Carbamyl-Lysine (CBL) Polyclonal Antibody

Immunoblot/ELISA

50 µg

STA-077

Rabbit Anti-Carbamyl-Lysine (CBL) Polyclonal Antibody

Immunoblot/ELISA

50 µg

STA-078

N/A

10 µg

STA-379

OxiSelect™ Protein Carbamylation Sandwich ELISA

Carbamyl Lysine-BSA

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145

METABOLISM RESEARCH

Alcohol Assays

Alcohol Assay Kits Our Alcohol Assay Kits provide a convenient platebased method to measure primary alcohols, including ethanol, in plasma, serum or saliva samples.* The kit uses an enzymatic oxidation reaction that produces hydrogen peroxide, which reacts with the provided probe. Kits are available with either colorimetric or fluorescence-based detection, both of which are performed in a 96-well microtiter plate. The colorimetric assay can detect alcohol levels as low as 30 ÂľM, while the fluorometric assay detects as low as 15 ÂľM. *These assays are not suitable for urine samples. Product Name Alcohol Assay Kit

Standard Curve Generated with the Alcohol Assay Kit. Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-620

Fluorometric

100 Assays

STA-621

Free Glycerol Assay Kits Glycerol is the backbone of triglycerides. Lipases secreted in the intestines hydrolyze the triglyceride ester bond, producing glycerol and free fatty acids. Our Free Glycerol Assay Kits use a coupled enzymatic reaction system to measure free, endogenous glycerol concentrations. The glycerol is phosphorylated and oxidized, producing hydrogen peroxide, which reacts with the probe provided with each kit. Kits are available with either colorimetric or fluorescence-based detection, both of which are performed in a 96-well microtiter plate. Product Name Free Glycerol Assay Kit

146

Phone 1-858-271-6500

Standard Curve Generated with the Free Glycerol Assay Kit.

Detection

Size

Catalog Number

Colorimetric

100 Assays

STA-398

Fluorometric

100 Assays

STA-399

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

PATHOGEN AND TOXIN ASSAYS

Viral Core Antigens

Virus Core Antigen Detection Core antigens provide a convenient way to detect and quantify certain infectious viruses. We offer a variety of ELISA kits to measure the core antigen levels in plasma, serum, or purified virus preps. Note: Assays are for research use only, not for clinical diagnostic use.  

MuLV Core Antigen HIV-1 p24 Core Protein

 

Hepatitis B Core Antigen Hepatitis C Core Antigen

QuickTiter™ MuLV Core Antigen ELISA Kit Murine leukemia virus (MuLV) is a retrovirus capable of causing cancer in mice and related vertebrates. Recently discovered in humans, xenotropic murine leukemia virus-related virus (XMRV) is closely related to MuLV; the p30 core antigens share 96% identity.

2.5

O D 450nm

Our QuickTiter™ MuLV Core Antigen ELISA Kit specifically measures the level of the p30 core antigen in blood or other samples.

2 1.5 1 0.5

 Sensitive: Detect as little as 300 pg/mL  Fully quantitative: Recombinant core antigen included as positive control

0 0

MuL V p30 (ng /mL ) Standard Curve Generated with the QuickTiter HBV Core Antigen ELISA Kit.

Product Name QuickTiter™ MuLV Core Antigen ELISA Kit (MuLV p30)

Detection

Size

Catalog Number

Colorimetric

96 Assays

VPK-156

QuickTiter™ HIV-1 p24 ELISA Kit A popular method of quantifying HIV-1 is the p24 ELISA. Our p24 ELISA kits provide a quick, convenient way to quantify the concentration of your lentivirus.

Product Name

Detection

QuickTiter™ HIV Lentivirus Quantitation Kit (HIV-1 p24 ELISA)

148

Phone 1-858-271-6500

Colorimetric

USA Toll-Free 1-888-CBL-0505

Size

Catalog Number

96 Assays

VPK-108-H

5 x 96 Assays

VPK-108-H-5

Fax 1-858-271-6514

Viral Core Antigens

PATHOGEN AND TOXIN ASSAYS

QuickTiter™ HBV Core Antigen ELISA Kit The QuickTiter™ HBV Core Antigen ELISA Kit specifically quantifies the core protein of Hepatitis B virus. A mouse monoclonal antibody is coated onto an 8 x 12 strip-well plate which allows the flexibility to save some of the wells for later use. The quantity of HBV is compared to the provided HBV Core Antigen Standard.

 Sensitive: Detect as little as 1 ng/mL  Fast: Results in about 5-6 hours  Fully Quantitative: Recombinant core antigen included as positive control

Product Name

QuickTiter HBV Core Antigen ELISA Kit Standard Curve.

Detection

QuickTiter™ HBV Core Antigen ELISA Kit

Colorimetric

Size

Catalog Number

96 Assays

VPK-150

5 x 96 Assays

VPK-150-5

QuickTiter™ HCV Core Antigen ELISA Kit The QuickTiter™ HBV Core Antigen ELISA Kit specifically quantifies the core protein of Hepatitis B virus. A mouse monoclonal antibody is coated onto an 8 x 12 strip-well plate which allows the flexibility to save some of the wells for later use. The quantity of HBV is compared to the provided HBV Core Antigen Standard.

 Sensitive: Detect as little as 1 ng/mL  Fast: Results in about 5-6 hours  Fully Quantitative: Recombinant core antigen included as positive control

Product Name QuickTiter™ HCV Core Antigen ELISA Kit

QuickTiter HCV Core Antigen ELISA Kit Standard Curve.

Detection

Size

Catalog Number

Colorimetric

96 Assays

VPK-151

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149

PATHOGEN AND TOXIN ASSAYS

Toxin Assays

OxiSelect™ BPDE Adduct ELISA Kits Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, cigarette smoke, and automotive exhaust. Benzo(a)pyrene, the first chemical carcinogen discovered, is converted to benzo(a)pyrene 7,8 diol-9,10 epoxide (BPDE) through a series of enzymatic reactions. BPDE can attack both proteins and DNA, forming adducts that can potentially result in tumor formation. Our OxiSelect™ BPDE Adduct ELISA Kits provide a convenient method to measure BPDE adducts with either DNA or proteins from cells or tissues. Quantitation is performed by comparing unknown values against a standard curve generated with the standard provided in each kit.  Our BPDE DNA Adduct ELISA can detect adducts as low as 30 ng/mL.  Our BPDE Protein Adduct ELISA can detect adducts as low as 60 ng/mL. Recent Product Citation Chiu, C.Y. et al. (2014). Low-dose benzo(a)pyrene and its epoxide metabolite inhibit myogenic differentiation in human skeletal muscle-derived progenitor cells. Toxicol. Sci. 10.1093/toxsci/ kfu003. (STA-357) Product Name

Standard Curve Generated Using the OxiSelect™ BPDE DNA Adduct ELISA Kit.

Detection

Size

Catalog Number

OxiSelect™ BPDE DNA Adduct ELISA Kit

Colorimetric

96 Assays

STA-357

OxiSelect™ BPDE Protein Adduct ELISA Kit

Colorimetric

96 Assays

STA-301

Aflatoxin ELISA Kits Aflatoxins are among the most potent genotoxic agents known. They can induce chromosomal abberations, micronuclei, sister chromatid exchange, unscheduled DNA synthesis, and chromosomal strand breaks. Aflatoxins can form adducts with both DNA and proteins. Our Aflatoxin Competitive ELISA Kits provide a convenient method for detection of aflatoxin adducts.  Our Aflatoxin Competitive ELISA Kit measures the total of Aflatoxin B1 and Aflatoxin B2 adducts in protein samples.  Our Aflatoxin DNA Adduct Competitive ELISA Kit detects total Aflatoxin B1-DNA adducts, both ring-opened and ring-closed forms. Product Name

150

Standard Curve Generated with the Aflatoxin Competitive ELISA Kit. Detection

Size

Catalog Number

Aflatoxin Competitive ELISA Kit

Colorimetric

96 Assays

AKR-350

Aflatoxin DNA Adduct Competitive ELISA Kit

Colorimetric

96 Assays

AKR-351

Phone 1-858-271-6500

USA Toll-Free 1-888-CBL-0505

Fax 1-858-271-6514

PRODUCT INDEX Product 293 Cell Lines AAV Adenovirus GFP Stable Expression Lentivirus Retrovirus 3-Nitrotyrosine Antibodies ELISA Kit 4-HNE Antibodies ELISA Kit 6-4PP Quantitation Kits 8-Iso-Prostaglandin F2 ELISA 8-Isoprostane ELISA Kit 8-Nitroguanine ELISA Kit 8-OHdG ELISA Kit 8-OHG ELISA Kit A549/GFP Cell Line AAV (Adeno-Assoc. Virus) Cell Line Expression Systems Expression Vectors Helper Free Systems Packaging Systems Premade Control Viruses Purification Kits Quantitation Kit shRNA Expression Titer Kit Transduction Reagent Acetylcholine Assays Active Rac-GEF Assay Adenovirus Cell Line Expression Systems microRNA Expression Premade Recombinant Purification Kits Quantitation Kits RCA Assay shRNA Expression Titer Kits Transduction Reagent Adhesion Assays Adipogenesis Assay Advanced Glycation End Products Assay Advanced Oxidation Protein Products Assay Aflatoxin Assays AGE Assays Albumin Antibody ELISA Kit Protein

Page 47 50 123 59 67 85 85 92 92 98 92 92 95 94 95 123 47 41-45 46 41-45 45 46 47 48 41-45 48 48 140 114 50 49, 81 81 50-53 54 55 56 49 55 56 10-11 27 88-89 87 150 88-89 142 142 142

Product Alcohol Assays Aldehyde-Induced DNA Damage Assays Alkaline Phosphatase Assays Angiogenesis Recombinant Adenovirus Tube Formation Assay Anoikis Assays Antibodies Albumin Apolipoproteins Beta-Actin Beta-Tubulin Carboxymethyl Lysine (CML) C-Reactive Protein Flag Tag Fluorescent Proteins GAPDH GFP GST Tag HA Tag His Tag HNE (4-Hydroxynonenal) MDA (Malondialdehyde) Methylglyoxal (MG) Myc Tag Nitrotyrosine Plasminogen RFP Antibody Tools Blocking Reagent Purification Kit Western Stripping Solution Antioxidant Assays Catalase Activity Assay Cellular Antioxidant Asssay Glutathione Assay Glutathione Reductase Assay HORAC Assay ORAC Assay Superoxide Dismutase Activity Assay Total Antioxidant Capacity Assay AOPP Assay AP Sites Quantitation Kit Apo(a) ELISA ApoAI ELISA ApoAI/ApoB Duplex ELISA ApoAII ELISA ApoB ELISA ApoCI ELISA ApoCII ELISA ApoCIII ELISA ApoE ELISA

www.cellbiolabs.com

Page 146 99 38 50 29 23 142 133 125 125 89 142 125 124 125 124 125 125 125 92 91 89 125 85 142 124 126 125 126 106 109 108 108 110 110 107 110 87 96 132 132 133 132 132 132 132 132 132

Product Apolipoproteins Antibodies ELISA Kits Proteins Arf1 Activation Assay Arf6 Activation Assay AUF1 Retroviral Vector Autophagy Expression Vectors -Actin Antibody -Galactosidase Recombinant Adenovirus Reporter Assays ß-Tubulin Antibody Bacterial Protein Extraction Reagents Biochips for Cell Adhesion Blocking Reagent BPDE DNA Adduct ELISA Protein Adduct ELISA BrdU ELISA Kit BT-549/GFP Cell Line C3 Expression Vector CA9 Recombinant Adenovirus c-Abl Retroviral Vector cAMP ELISA Kits Cancer Cell Assays Angiogenesis Anoikis Cell Adhesion Cell Invasion Cell Migration Cell Transformation Colony Formation Soft Agar Tumor Cell Isolation Kit Tumor Sensitivity Carbamyl Lysine Antibodies ELISA Kits Carbonyl Assays Carboxyethyl Lysine ELISA Carboxymethyl Lysine Assays Catalase Activity Assays Cdc42

Page 133 132 132 112113 112113 68 30 125 50 123 125 127 10 126 150 150 25 123 116 50 67 119 29 23 10-11 18-19 12-19 6-7 6-9 6-9 9 8 145 145 86 88 89 106

Activation Assay

112-

Agarose Beads Recombinant Adenovirus Recombinant Protein Retroviral Vector

115 51 118 69

info@cellbiolabs.com

151

PRODUCT INDEX Product CEA Recombinant Adenovirus CEL ELISA Kit Cell-Based Assays Adhesion Angiogenesis Anoikis Cell Contraction Cell Viability Chemotaxis Colony Formation Cytotoxicity Haptotaxis Invasion Migration Phagocytosis Proliferation Senescence Soft Agar Transformation Transmigration Tumor Sensitivity Wound Healing Cell Cycle Adenoviruses Anoikis Assay Cell Viability Assay Cytotoxicity Assay Retroviral Vectors Senescence Assays Cell Fractionation Kits Cell Invasion Assays Cell Lines 293AAV 293AD 293LTV 293RTV 293/CFP 293/GFP 293/Luc 293/YFP 293T/GFP-Puro A549/GFP BT-549/GFP ES-2/GFP HeLa/GFP JK1 Feeder Cells MCF-7/GFP MCF-7/Luc MDA-MB-231/GFP MDA-MB-231/GFP-RFP MDA-MB-231/Luc MDA-MB-231/RFP MDA-MB-436/GFP MDA-MB-436/RFP MDA-MB-468/GFP

152

Page 50 88 10-11 29 23 29 22 15, 19 6-9 22 16 18-19 12-19 28 24-25 23 6-9 6-7 17 8 20 51 23 22 22 67 23 127 18-19 47 50 59 67 123 123 123 123 123 123 123 123 123 35 123 123 123 123 123 123 123 123 123

Phone 1-858-271-6500

Product Cell Lines (cont’d) MEF Feeder Cells NIH3T3/GFP OVCA429/GFP OVCAR-5/RFP Plat-A Retroviral Packaging Cells Plat-E Retroviral Packaging Cells Plat-GP Retroviral Packaging Cells SKOV-3/GFP-Luc SKOV-3/Luc SKOV-3/RFP SNL Feeder Cells T47D/GFP Cell Migration Assays Cell Proliferation Assays Cell Transformation Assays Cell Viability Assay Cellular Antioxidant Assay Cellular Senescence Assays CETP ELISA Kit cGMP ELISA Kits Checkpoint Kinase Assays Chemotaxis Assays Cholesterol Assays Cholesteryl Ester Transfer Protein Assay Clonogenic Tumor Cell Isolation Kit CML (Carboxymethyl Lysine) Antibodies Assays CML-LDL ELISA Kit c-Myc Retroviral Vectors Colony Formation Assays Cell Transformation Assays Hematopoietic Colony Forming Cell Assay Stem Cell Colony Assay Tumor Cell Isolation Kit Tumor Sensitivity Assay Comet Assay Kits & Slides Core Antigen Assays CPD Quantitation Kits C-Reactive Protein Antibody ELISA Protein Cre Recombinant Adenovirus Creatinine Assay CSK Recombinant Adenovirus

Page 35 123 123 123 64 64 64 123 123 123 35 123 12-19 24-25 6-7 22 109 23 136 119 121 15, 19 130131 136 9 89 89 93, 134 69 6-7 36 37 9 8 97 14898 142 141 142 50 145 53

USA Toll-Free 1-888-CBL-0505

Product Cyclic AMP ELISA Kits Cyclic GMP ELISA Kits CytoSelect™ Cell-Based Assays Anoikis Cell Adhesion Cell Invasion Cell Migration Cell Transformation Cell Viability Chemotaxis Colony Formation Cytotoxicity Haptotaxis Phagocytosis Soft Agar Transmigration Tumor Sensitivity Wound Healing Cytoskeleton Regulation Activation Assays

Page 119 119

23 10-11 18-19 12-19 6-7 22 15, 19 6-9 22 16 28 6-9 17 8 20 112114 51 116 69 22

Adenoviruses Expression Vectors Retroviral Vectors Cytotoxicity Assay DCC Recombinant 51 Adenovirus DNA Damage Assays 6-4PP Quantitation 98 8-Nitroguanine ELISA Kit 95 8-OHdG ELISA Kit 94 Aldehyde-Induced Damage 99 AP Sites Quantitation Kit 96 BPDE Adduct ELISA 99 Comet Assays 97 CPD Quantitation 98 Double-Strand Break Assay 96 UV Damage 98 DNA Methylation Assays 100 ECM (Extracellular Matrix) Assays Cell Adhesion Assays 10 Cell Invasion Assays 18-19 Tube Formation Assay 29 Endothelial Tube Assay 29 Epitope Tag Antibodies 125 ERK2 Recombinant Adenovirus 52 Retroviral Vector 68 ERK5 Recombinant 52 Adenovirus ES/EC Cells Alkaline Phosphatase Assays 38 Colony Formation Assays 37 Retroviral Expression 34 Systems

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PRODUCT INDEX Product ES-2/GFP Cell Line Ethanol Assays Exoenzyme C3 Expression Vector Extracellular Matrix Kits Cell Adhesion Assays Cell Invasion Assays Tube Formation Assay Feeder Cells FFA Assay Kits Firefly Luciferase Recombinant Adenovirus Flag Tag Antibody Free Fatty Acid Assays Free Glycerol Assays Fyn Recombinant Adenovirus Gap Closure Migration Assays GAPDH Antibody GEF (Guanine Exchange Factors) Activation Assays Agarose Beads GFP Antibody ELISA Kit Lentiviral Vectors Quantitation Kits Recombinant Adenovirus Recombinant Lentivirus Recombinant Protein Retroviral Vectors Stable Cell Lines GGA3 Agarose Beads Global DNA Methylation Assays Glutathione Assay Glutathione Reductase Assay Glycerol Assays Glycoaldehyde-BSA GPCR Signaling Products GST Antibody Inclusion Body Solubilization and Renaturation Kit GTPase Assay Kits HA Tag Antibody Haptotaxis Assays HBV Core Antigen ELISA HCV Core Antigen ELISA HDL Assay Lipoprotein, Human HEK 293 Cell Lines AAV Adenovirus

Page 123 146 116 10 18-19 29 35 139 50 125 139 146 53 13 125

114 115 124 122 59 122 50 59 124 67 123 115 100 108 108 146 88 119 125 127 112113 125 16 149 149 131 131 47 50

Product HEK 293 Cell Lines (cont’d) GFP Stable Expression Lentivirus Retrovirus HeLa/GFP Cell Line Hematopoietic Colony Forming Cell Assay Hepatitis B Core Antigen ELISA Hepatitis C Core Antigen ELISA HIF-1

Page 123 59 67 123 36 149 149 26, 101 50

Assays Recombinant Adenovirus High Density Lipoprotein Assay Protein His Tag Antibody Protein ELISA HIV-1 p24 ELISA Kits

131 131 125 125 60-61, 148

HNE Antibodies Assays HNE-LDL Assay hnRNPA0 Retroviral Vector HORAC Assay Kit H-Ras Activation Assay Recombinant Protein HuB Retroviral Vector HuC Retroviral Vector HuD Retroviral Vector HuR Retroviral Vector Hydrogen Peroxide Assays Hydroxyl Radical Antioxidant Capacity Assay Hypoxia Assays

92 92 93, 134 68 110 112113 118 68 68 68 68 104 110 26,

IFN Recombinant Adenovirus 52 IB Recombinant Adenovirus 53 IKK Recombinant Adenovirus 53 IL-2 Recombinant Adenovirus 52 Immunoblot Blocking 126 Reagent In Vitro Angiogenesis Assay 29 In Vitro Tumor Sensitivity 8 Assay Inclusion Body Solubilization 127 Induced Pluripotent Stem Cells Lentiviral Vectors 33 Retroviral Packaging Cells 32 Retroviral Vectors 32-33

www.cellbiolabs.com

Product Invasion Assays iPS Cell Reprogramming Lentiviral Vectors Retroviral Packaging Cells Retroviral Vectors JK1 Feeder Cells JNK1 Recombinant Adenovirus Retroviral Vector Klf4 Retroviral Vectors KOSM Viral Vectors K-Ras Activation Assay Recombinant Protein LC3 Expression Vectors LCAT Assays LDH Cytotoxicity Assay LDL Assays Lipoprotein, Human Oxidized LDL Receptor Assay Lecithin Cholesterol Acyltransferase Assays Lentivirus Cell Line Concentration & Purification Kits Control Plasmids Expression Systems Expression Vectors Packaging Systems Premade Control Viruses Purification Kits Quantitation Kits shRNA Expression Titer Kits Transduction Kits Leukocyte Assays Adhesion Transmigration Lin-28 Retroviral Vectors Lipid Extraction Kit Lipid Peroxidation Assays 8-Isoprostane ELISA HNE Adduct ELISA Malondialdehyde (MDA) Assays TBARS Assay Lipid Quantification Assay Lipoprotein Lipase Assays Lipoproteins Assays Human Oxidized

info@cellbiolabs.com

Page 18-19 33 32 32-33 35 52 68 69 33 112113 118 30 137 22 131 131 134 135 137 59 62 59 57-58 59 58 59 62 60-61 57-58 60-61 63 11 17 69 143 92 92 90-91 90 143 138 130133 131132 134

153

PRODUCT INDEX Product LOX-1 Assay LPL Assays Luciferase Recombinant Adenovirus Reporter Cell Lines Malondialdehyde Antibodies Assays MAP Kinase Signaling Recombinant Adenovirus Retroviral Vectors MAPKAPK2 Recombinant Adenovirus Retroviral Vector MCF-7/GFP Cell Line MCF-7/Luc Cell Line MDA (Malondialdehyde) Antibodies Assays MDA-LDL Assay MDA-MB-231/GFP Cell Line MDA-MB-231/Luc Cell Line MDA-MB-231/RFP Cell Line MDA-MB-436/GFP Cell Line MDA-MB-436/RFP Cell Line MDA-MB-468/GFP Cell Line MEF Feeder Cells MEK1 Recombinant Adenovirus Retroviral Vector

Page 135 138 50 123 91 90-91 52 68 52 68 123 123 91 90-91 93, 134 123 123 123 123 123 123 35 52 68

MEK5 Recombinant MEKK1 Recombinant Adenovirus MEKK3 Recombinant Adenovirus Methylglyoxal (MG) Antibody ELISA Kits Microfluidic Biochips microRNA Analysis Adenoviral Expression System Clone Collection Control Vectors Expression Vectors Functional Reporter System Precursor Clone Collection Retroviral Expression Vector Transduction Enhancer Migration Assays miRNA Analysis Adenoviral Expression System Clone Collection Control Vectors

154

52 52 52 89 89 10 81 74-80 80 74-80 82 74-80 81 82 12-19 81 74-80 80

Phone 1-858-271-6500

Product miRNA Analysis (cont’d) Expression Vectors Functional Reporter System Precursor Clone Collection Retroviral Expression Vector Transduction Enhancer MKK3 Recombinant Adenovirus Retroviral Vector MKK4 Recombinant Adenovirus MKK6 Recombinant Adenovirus Retroviral Vector MKK7 Recombinant Adenovirus MTT Cell Proliferation Assay MuLV p30 Core Antigen ELISA Myc Tag Antibody MyoD Recombinant Adenovirus Myogenin Recombinant Adenovirus myr-Akt Retroviral Vectors myr-Rac1 Recombinant Adenovirus Retroviral Vectors NANOG Retroviral Vectors N-Carboxyethyl Lysine ELISA N-Carboxymethyl Lysine Antibody Assay Kits NFB Recombinant Adenoviruses NIH3T3/GFP Cell Line Nitrated LDL Nitric Oxide Assays Nitroguanine ELISA Kit Nitrotyrosine Antibodies Assay Kits NOD2 Recombinant Adenovirus N-Ras Activation Assay Recombinant Protein Nuclear / Cytosolic Cell Fractionation Kits NY-ESO-1 Recombinant Adenovirus Oct-3/4 Retroviral Vectors OHdG ELISA Kit OHG ELISA Kit

Page 74-80 82 74-80 81 82 52 68

Product ORAC Assay Kit OVCA429/GFP Cell Line OVCAR-5/RFP Cell Line Oxidized LDL Assay Kits Lipoprotein, Human Oxidized Proteins OxiSelect™ Oxidative Stress / Damage Assays

Page 110 123 123 93, 134 131 89

Lipid Peroxidation Assays Protein Oxidation Assays

106110 94100 90-93 85-89

ROS Assays

102-

148

OxLDL Assay

93,

125

OxPL Assay Oxygen Radical Antioxidant Capacity (ORAC) Assay

134

Antioxidant Assays

52 68

DNA / RNA Damage Kits

51 51

p24 ELISA Kits

p38 Recombinant Adenovirus Retroviral Vectors p53 Recombinant Adenovirus Retroviral Vectors p68 RNA Helicase Adenovirus PABP Retroviral Vector PAK1 PBD Agarose Beads Recombinant Adenovirus PCNA ELISA Kit PCSK9 ELISA Kit Peroxide Detection Assays Phagocytosis Assays Phosphatidylcholine Assay Phospho Antibody Stripping Solution PhosphoBLOCKER™ Western Blot Blocking Reagent Phosphoproteins Antibody Stripping Solution Blocking Reagent Purification Kit PI3K Retroviral Vector PKC Recombinant Adenovirus Plasminogen Antibody ELISA Kit Protein, Human

52 69 69 88 89 89 53 123 131 105 95 85 85 53 112113 118 127 50 69 94 95

USA Toll-Free 1-888-CBL-0505

110 60-61, 148 52 68 51 67 51 68 115 51 25 136 104 28 140 126 126

126 126 126 68 53 142 141 142

Fax 1-858-271-6514

PRODUCT INDEX Product Plat-A Retroviral Packaging Cells Plat-E Retroviral Packaging Cells Plat-GP Retroviral Packaging Cells Platinum Retroviral Expression Expression Systems Packaging Cell Lines pMX Retroviral Vectors PRAK Recombinant Adenovirus Retroviral Vector Proliferating Cell Nuclear Antigen Assay Proliferation Assays Protease Retroviral Vectors Protein Extraction Reagents Protein Oxidation Assays Advanced Glycation End Products (AGE) Advanced Oxidation Protein Products (AOPP) BPDE Adduct Carbonyl CEL (Carboxyethyl Lysine) CML (Carboxymethyl Lysine) MG (Methylglyoxal) Nitrotyrosine Protein Phosphorylation Antibody Stripping Solution Blocking Reagent Purification Kit Protein Quantitation Kit Proteins Albumin Apolipoproteins EGFP GRP-PH Domain Oxidized/Nitrated Purification Kits AAV Adenovirus Antibodies Lentivirus Phosphoproteins Retrovirus QuickTiter™ Viral Titer & Quantitation Kits AAV Adenovirus HBV Core Antigen HCV Core Antigen HIV p24 Lentivirus, Recombinant

Page 64 64 64

Product

Page

QuickTiter™ Viral Titer & MuLV p30 Retrovirus, Recombinant Rab Recombinant Proteins Rac Activation Assays

65 64 66-69 52 68 25 24-25 69 127

Agarose Beads GEF Assay Recombinant Adenovirus Recombinant Proteins Retroviral Vectors Radius™ Cell Migration Assays Raf1 Recombinant Adenovirus Retroviral Vectors Ral

88-89 87 87 86 88 89 89 85

142 132 124 119 131

112113 118

47 56 125 62 126 70

48 55 149 149 148 60-61

Recombinant Proteins RAPAd® Adenoviral Expression Systems Rapid GST Inclusion Body Solubilization and Renaturation Kit Rapid RCA Assay Ras Superfamily Activation Assays Agarose Beads Expression Vectors Recombinant Adenovirus Recombinant Proteins Retroviral Vectors RCA Assay Kit Reactive Oxygen Species (ROS) Assays Recombinant Adenoviruses Recombinant Proteins Fluorescent Proteins GRP-PH Domain Small GTPase Rel B Recombinant Adenovirus

Quantitation Assays

Activation Assays

Activation Assays 126 126 126 128

Recombinant Adenovirus Recombinant Lentivirus Retroviral Vectors Stable Cell Lines Retrovirus Concentration & Purification Kits Expression Systems Expression Vectors Gene-Specific Vectors Packaging Cell Lines Purification Kits Quantitation Kits shRNA Expression Transduction Kits RFP Antibody ELISA Kit Recombinant Protein Stable Cell Lines RGS Recombinant Proteins Rho

112113 115 118

Agarose Beads Recombinant Protein Rap

144-

112113 115 114 51 118 69

112113 115 118

Activation Assay

Renal Function Assays Reporter Genes Lentiviral Vectors

52 68

Agarose Beads Recombinant Proteins Ran

Page

148 71 118

Activation Assay

Product

Agarose Beads Recombinant Adenovirus Recombinant Proteins Retroviral Vector RhoA Activation Assay

127 RhoB Activation Assay 56 RhoC Activation Assay 112113 115 116 51 118 69 56 102105 50-53

www.cellbiolabs.com

124 119 118 53

Rho Kinase Activity Assays RIPA Buffer RNA Damage ELISA Kit RNAi Enhancer Reagent ROCK Activity Assay Kits ROS Assays scAAV Control Vector Expression Systems Expression Vector SCGE Assay Kits SEAP Recombinant Adenovirus Senescence Assays shAkt Recombinant Adenovirus

info@cellbiolabs.com

59 122123 50 59 67 123 70 65 66 67-69 64, 67 70 71 66 72 124 123 124 123 118 112113 115 51 118 69 112113 112113 112113 120 128 95 82 120 102105 46 42-45 46 97 50 23 53

155

PRODUCT INDEX Product shRNA Expression Adeno-Associated Virus Adenovirus Lentivirus Retrovirus Single Cell Gel Electrophoresis Assays SKOV-3/GFP-Luc Cell Line SKOV-3/Luc Cell Line SKOV-3/RFP Cell Line Small GTPase

Page 41-45 49 57-59 66 97 123 123 123

Activation Assays

112-

Active GEF Assays Agarose Beads Expression Vectors Premade Adenoviruses Retroviral Vectors SNL Feeder Cells SOD Activity Assay Kit Soft Agar Colony Assay Kits Cell Transformation Assays

114 115 116 51 69 35 107 6-7

Hematopoietic Colony

Stem Cell Colony Formation

Tumor Cell Isolation Kit Tumor Sensitivity Assay

9 8

SOK Recombinant

Sox2 Retroviral Vector Sphingomyelin Assay Src Recombinant Adenovirus Stat5 Retroviral Vectors Stem Cell Research Alkaline Phosphatase Detection Kits Feeder Cells

69 140 53 68 38 35

Hematopoietic Colony iPS Cell Reprogramming PCR Primers

36 32-33 38

Retroviral Expression Stem Cell Colony Assay Total Protein: ES Cell Line D3 Total RNA: ES Cell Line D3 Superoxide Dismutase Assay T47D/GFP Cell Line TAC Assay Tac-Rac1 Recombinant Adenovirus TBARS Assay Kit TIA1 Retroviral Vector Tiam1 Assay Kit TIAR Retroviral Vector

156

Phone 1-858-271-6500

34 37 38 38 107 123 110 52 90 68 114 68

Product Total Antioxidant Capacity Total Cholesterol Assays Total Protein: ES Cell Line D3 Total RNA: ES Cell Line D3 Transcription Regulation Retroviral Vectors Transformation Assays Transmigration Assays Triglyceride Assays TTP Retroviral Vector Tube Formation Assay Tumor Antigen Adenoviruses Tumor Cell Assays Cell Adhesion Cell Invasion Cell Migration Cell Transformation Chemosensitivity Soft Agar Colony Formation Transmigration Tumor Cell Isolation Tyrosine Kinase Adenoviruses uPA / uPAR Retroviral Vectors Urea Assay Uric Acid / Uricase Asay UV DNA Damage Assays V5 Tag Antibody VEGF Recombinant Adenovirus Very Low Density Lipoprotein Assay Lipoprotein, Human ViraBind™ Purification Kits AAV Adenovirus Lentivirus Retrovirus ViraDuctin™ Transduction Kits & Reagents AAV Adenovirus Lentivirus Retrovirus Viral Expression Systems AAV Adenovirus Lentivirus Retrovirus Viral Packaging Cells AAV Adenovirus Lentivirus Retrovirus

Page 110 130 38 38 68 6-7 17 139 68 29 50 10-11 18-19 12-19 6-7 8 6-9 17 9 53 69 144 144 98 125 50 131 131 47 54 62 70

Product Viral Titer Kits AAV Adenovirus Lentivirus Retrovirus Viral Transduction Reagents AAV Adenovirus Lentivirus Retrovirus ViraSafe™ Lentivirus Expression Systems Virus Core Antigen Assays HBVcAg HCVcAg HIV-1 p24 MuLV p30 Virus Purification Kits AAV Adenovirus Lentivirus Retrovirus Virus Quantitation Kits AAV Adenovirus Lentivirus Retrovirus VLDL Assay Lipoprotein, Human VSV-G Retroviral Vector Western Blot Blocking Reagent Wound Healing Assay WST-1 Cell Proliferation Reagent

Page 48 55 60-61 71 48 56 63 72 57-58 149 149 148 148 47 54 62 70 48 55 60-61 71

48 56 63 72 41-45 49, 81 57-58 65 47 50 59 64, 67

USA Toll-Free 1-888-CBL-0505

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131 131 67 126 20 24

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