Technology Overview
TARGATT™ Fast & Site-Specific Knock-in Technology
TARGATT™ Services and Products
Background:
Fast & Site-Specific Knock-in Mouse Service
Applied Stem Cell Inc.’s proprietary TARGATT™ technology enables highly efficient site-specific gene integration in mammalian cells and animals1, 2. This technology uses ФC31 integrase to insert any gene of interest into a docking site, pre-engineered in an intergenic region and transcriptionally active genomic locus. Our TARGATT™ technology improves several aspects in the generation of transgenic cell lines and animals: (1) High integration efficiency mediated by ФC31 integrase reduces time and cost; (2) Site-specific integration at a pre-selected genomic locus eliminates position effect and ensures high level expression of the transgene; (3) Integration at intergenic region ensures that no internal genes are interrupted; (4) Single copy gene integration eliminates repeat-induced gene silencing and genomic instability; (5) Site-specific integration allows a precise comparison of the effects of the transgenes among different lines. TARGATT™ technology can be utilized for a variety of applications including reporter gene expression, gene knockdown, disease cell and animal models.
in vitro Model
Knock-in Mouse Generation
Stable Cell Line Generation
1 Embryos with attp sites are collected
1 Generate master TARGATT™ Cells
from TARGATT™ mouse.
2
attB 3
TARGATT™ embryos.
4 Screening pups for site-specific gene integration
4
Transfection
Knock-in Mouse Service (C57BL/6 strain) Knock-in Mouse Service (FVB strain)
Comparison of timelines for Knock-in Mouse Models TARGATT™ System Vector Design
Vector Design
Pronuclear Microinjection
ES cell Targeting
F0 Genotyping
3 months
Reliable Gene Expression
AST-5021 AST-5001
Traditional Knock-in
$19,000 $17,000
vs Blastocyst microinjection
Chimeras
Germline Mice
10 months
Knock-in Cell Line within 1 - 3 months
TARGATT™ Embryos & Transgenic Kit If you prefer to generate a knock-in mouse model on your own, TARGATT™ Embryo and Transgenic Kits are available for purchase. For your convenience, you may also mix and match any of our TARGATT™ products and services.
Applications
in vitro screenings Drug discovery Study drug interactions Toxicity Study Personalized medicine
in vivo screenings Generation of humanized models Generation of disease models Generation of drug / genome interaction models
Features: Fast: Our proprietary technology will generate your site-specific
TARGATT™ Cell Lines Expressing Gene of Interest
Knock-in Mouse within 3 months
Applications
Low Yes Yes One copy 10 Months
Vector design and construction Integration enzyme generation DNA preparation Embryo microinjection / Implantation Animal care-housing Genotyping
TARGATT™ Vector with Gene of interest (eg. GFP)
3 Pronuclear injection into
Up to 40% Yes Yes One copy 3 Months
Service Includes:
with attp sites (6 months)
2
Efficiency Site-Specificity Transgene Expression Transgene Copy Number Turnaround Time
Traditional Rosa26 Knock-in
knock-in mouse in 3 months. Reliable: Any gene of interest can be specifically inserted at a defined, transcriptionally active locus. We guarantee transgene expression. Versatile: Tissue specific and/or ubiquitous expression are available.
How it works: in vivo Model
TARGATT™ System
TARGATT™ H11 Embryos FVB (Frozen) TARGATT™ H11 Embryos C57BL/6 (Frozen) TARGATT™ Rosa26 Embryos FVB (Frozen) TARGATT™ Transgenic Kit
References: 1. B. Tasic et al., Proc Natl Acad Sci USA. 108, 7902 (2011) . 2. J. Rossant, L. M. J. Nutter, M. Gertsenstein, Proc Natl Acad Sci USA. 108, 7659
AST-0002 AST-0012 AST-0004 AST-1002
$3,600* $4,800* $3,600* 2 rounds of microinjections $750
MTA is required upon ordering. *Quotes for academic research. Please contact us for industrial rates.
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Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Other Mouse Models / Gene Targeting Services
Cell Line Gene Modification Service
Conventional Knock-in/out Mouse Generation Service
Based on our proprietary site-specific TARGATT™ technology for fast knock-in (KI) mouse generation, Applied StemCell (ASC) recently developed a TARGATT™ system specifically for introducing gene(s) of interest into mammalian cell lines. The unique advantage of this system is that any gene of interest can be inserted into a defined, transcriptionally-active locus with high gene expression in a single copy fashion.
Technical Advantages of TARGATT™: TARGATT™ System
Homologous Recombination
Transposon
ZFN
Yes
Yes
Yes
No
No
Knockin efficiency
High
Very low
Low
High
High
Transgene copy
Single
Single
Single
Multiple
Multiple
Off-target event
Very Low
Very Low
Yes
Yes
Yes
High Expression
Time Consuming
Costly & High mutation rates
Insertional Mutations
Biosafety Level 2
Service Includes:
Vector construction Transfection / selection Cell screening Cell expansion / validation Cell cryopreservation
Knock-in/out Mouse Generation (C57BL/6 strain)
ASC-5023 $30,000
Knock-in/out Mouse Generation (129 / hybrid)
ASC-5003 $26,000
Behavioral Phenotyping Service for Knock-in/out Mice Compare behavioral differences between transgenic mice & wild type mice.
TARGATT™ Knock-in Cell Line Service AST-6001 $25,000 Custom Knock-in Cell Line Service AST-6021 Inquire
Molecular Cloning Services
Master Cell Line
Cell Lines Expressing Gene of Interest
DNA Cloning Services Examples of services:
Stable Cell Line Generation TARGATT™ Vector with Gene of Interest (eg. GFP) attB
Characterizaton (RT-PCR, Staining)
Cloning of amplified DNA fragments Gene targeting vector construction for homologous recombination Bacterial Artificial Chromosome (BAC) recombineering Generation of RNAi and inducible vectors Site-specific mutagenesis De novo gene synthesis Knock-in/out Vector ASC-7001 $6,000 - $9,000 Gene Expression Vector ASC-7002 $3,000 - $6,000
3 months
Virus Packaging Service Stem Cell Differentiation and Characterizaton
Blood Cell
Gene targeting vector construction ES cell targeting Cre/FLP recombination Chimera production via blastocyst injection Chimera breeding to germline Mouse genotyping
Adding TARGATT™ sites to your cell line allows you to insert any gene of interest in a specific locus in the genome.
Application of the TARGATT™ System for the Generation of Site-Specific Knock-in Mammalian Cell Lines
Cells with attp site
Service Includes:
Chimera from blastocyst injection
Viral System
Site-Specific
Others
ASC provides high quality, one stop services for the generation of gene targeted mouse models. Cost-effective: Applied StemCell, Inc. has years of experience and a high success rate in generating custom gene targeted mice using our proprietary germline transmittable mESC lines. High Quality: We have high QC standard checkpoints at every step to ensure your project progresses smoothly. Fast Turnaround Time: We streamline every step. Gene targeted mouse models can be generated in as fast as 7 months.
Neuron
Promo
We provide high titer to ultra-high titer, ready to use VSV-G pseudoviral packaging services. Lentivirus Packaging Service Lentivirus Packaging Service
Regular Titer High Titer
>10^7 IFU/ml >10^8 IFU/ml
ASC-7011 $900 ASC-7012 $1,100
Lentivirus Packaging Service
Ultra-High Titer >10^9 IFU/ml
ASC-7013 $1,400
Retrovirus Packaging Service Regular Titer Retrovirus Packaging Service Regular Titer
Other Cell Types
>10^6 IFU/ml >10^7 IFU/ml
ASC-7021 $1,000 ASC-7022 $1,200
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Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Derivation and Characterization Services
Teratoma Formation Analysis Service Features:
To publish your new ES or iPS cell line, do you need: - Stringent molecular and functional assays to evaluate pluripotency criteria? - Karyotype analysis to rule out genetic aberrations?
- Turnaround time: 4- 12 weeks - Number of cells required / injection site: Mouse ESCs/iPSCs = 0.5 x 10^6 Human ESCs/iPSCs = 1 x 10^6 - Success Rate: >97%
We offer all services needed for a complete characterization of your pluripotent cell lines.
Service Highlights:
Derivation Services Derivation
Mouse ESC Derivation Service
ASC-6001
$2,900
Injection Specifics: A total of 4 mice (3 experimental and 1 control) will be used. Each recipient mouse will be injected at two locations: in the kidney capsule and in the testis. Steps: Cell injection, teratoma harvesting, Tissue Sectioning, H&E staining and histology analysis of teratoma sections.
Success Rate
Our scientists have successfully derived over 80 new mouse embryonic stem cell lines, including those from gene targeted embryos.
Key points
Characterization Services
Pluripotency Characterization
(P.9)
Immunocytochemistry (Mouse) ASC-6006 Immunocytochemistry (Human) ASC-6007 RT-PCR Inquire EB Formation Service (P.8) ASC-9020 Teratoma Formation Analysis (P.6-7) ASC-6008
Germline Transmission
Karyotyping
Chromosome Counting G-banding (Mouse) G-banding (Human)
$599
hESC
hiPSC
hESC
Teratoma Formation Rate
100%
100%
93.7%
25-40%
Differentiation
distinctive
distinctive
distinctive
poor
Turnaround Time
3-5 weeks
5-8 weeks
5-10 weeks
12-18 weeks
Cells needed
0.5-1 million/site
0.5-2 million/site
1-2 million/site
3-5 million/site
$799
Service Includes: $980 $2,900
$400
ASC-6003-GM $700 ASC-6003-GH
$700
Detection of genetic abnormalities Chromosome counting of metaphase spreads and G-banding
Sex Determination
ASC-6004
$200
Is it male or female? We will give you the answer!
ASC-6008
$2,900
References: K. Chen et al., Stem Cell Research USA. 9, 237–248 (2012) We are the exclusive teratoma service provider for Coriell Institute (Patient Specific iPSCs). See our service reports here: http://ccr.coriell.org/Sections/Collections/NIGMS/ipsc_list.aspx?PgId=696&coll=GM FAQ Q: What is your success rate for teratoma formation? A: After analyzing control and test samples, we have determined that 90% of our injections give rise to verifiable teratomas. Injections into both the kidney capsule and the testis ensures teratoma formation from pluripotent cells in one or both organs. The site and degree of teratoma formation is cell line dependent. For example, only 82% of hIPSC form teratomas in both the kidney and the testis. Other cell lines form teratomas in only the kidney or testis. By injecting three mice at two sites per test sample, we ensure that the best conditions are used to observe teratoma formation. Q: How long does it take to get results? A: To form mature teratomas, it takes 3-6 weeks for mouse ES/iPSCs and 5-10 weeks for hES/iPSCs. As a general practice, we allow the teratomas to grow for up to 12 weeks before we sacrifice the mice. Q: How do I prepare cells for the assay? A: Follow the instructions outlined in the Requisition Form found at http://www.appliedstemcell.com/userfiles/files/Teratoma_Service_Requisition_Form.pdf Typically, two T75 flasks are sufficient for the assay. We recommend that the cells be shipped (by one day delivery) at 70% confluence in flasks filled completely with culture media.
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Applied StemCell
1. Complete report with histological analysis and high quality/publication grade images of endoderm, mesoderm and ectoderm formation. 2. Tissue blocks and H&E stained tissue section slides Teratoma Formation Analysis (Human, Mouse)
ASC-6003-C
Traditional methods
mESC/miPSC
Type
iPSC Generation Service
ASC’s methods
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
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www.appliedstemcell.com
Embryoid Body (EB) Formation and Characterization Service
Technique Highlights Enhanced differentiation capability: Our technique of injection under both the kidney capsule and testis results in distinct teratoma formations with differentiated tissues derived from all three germ layers (A to F).
Features:
- A complete service that includes embryoid body (EB) formation and characterization to assess the pluripotency of your ESC/iPSC lines. - A fast alternative to traditional teratoma formation methods. Our turnaround time is approximately 3 weeks. - Ideal for rapid initial screenings or large sample screenings.
Service Includes: A
B
- Formation of EBs. - IHC staining of EBs for pluripotency markers (Alpha fetoprotein for endoderm, smooth muscle actin for mesodern and Beta III tubulin for ectoderm). - A full report that includes high resolution, publication-grade images.
C
EB Formation and Characterization Service ASC-9020 $980/sample (Additional $500 culture fee for frozen samples may apply) E
D
F
H
G
I
A : Cartilage B : Neuronal rosette C : Glands D : Pigmentaed cells E : Bone F : Stratified squamous epithelium G - I : Subcutaceous injections give rise to poorly differentiation tumor tissue.
I A
II
β-Tubulin III
SMA
High Teratoma Formation Rate: Three recipient mice per sample are dually injected under BOTH the kidney capsule and testis to ensure teratoma formation from pluripotent cells at one or both organs. ~82% of human iPS cell lines form teratomas in both the kidney and testis. 8.9% forms kidney tumor only, 2.6% forms testis tumor only. Kidney
Testis
+ + -
+ + -
B
AFP
% of iPSC Lines Producing Publication Grade Teratomas* 82.3 % 8.9 % 2.6 %
I. EB derived from GFP-expressing human ESCs.
6.2 %
II. β- Tubulin III (ecdoderm) SMA: Smooth muscle actin (mesoderm) AFP: Alpha fetal protein (endoderm)
*Teratomas with distinct tissue types
Satisfaction Guaranteed!! Please contact us with the information about your samples for details and pricing
Satisfaction Guaranteed!! Contact us with information about your samples for details and pricing 8
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Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Generation and Differentiation Services Genome Footprint-Free iPS Cell Generation Service
(mRNA/Episomal)
Frustrated with the amount of time and effort required to generate and characterize iPS cell lines? Wishing your valuable time was directed towards addressing the high priority issues of your research project rather than the time consuming tasks of iPS cell line generation and characterization? Let Applied StemCell generate the iPS cell line for you.
6-Step iPSC Generation Service can generate iPS cell lines using two proven genome footprint-free methods (mRNA or episomal vectors)
Cell Recovery and Quarantine Transfection with Episomal DNA Vectors or mRNA
Reprogramming Plasmids
Primary Cell Line (i.e. skin fibroblasts)
or
Colony Formation Colony Expansion Characterization via antibody staining and RT-PCR
hESC/iPSC Neuronal Differentiaton Service Do you want to differentiate your pluripotent cell line into neurons or glial cells? Let us make your life easier! We will make Neural Stem Cells (NSCs) or Neural Crest Stem Cells (NCSCs) for you! -You send us your pluripotent cell line (or we can make one for you) -We use our proprietary neural induction protocol and reagents to differentiate your ESC/iPSC into neural stem cells (NSCs) or neural crest stem cells (NCSCs) -NSCs and NCSCs are proliferating cells that can be frozen and cultured over a prolonged period of time. They are also capable of further differentiation into multiple neural cell types. NSCs Differentiation Service NCSCs Differentiation Service
ASC-8001 ASC-8001C
$9,500 $9,500
Related Services: Cardiomyocytes Differentiation Service Skeltal Muscle Differentiation Service
ASC-8003 ASC-8005
$5,000 $5,000 Neural Stem Cells (NSCs) / Neural Crest Stem Cells (NCSCs)
Primary Cell Line (i.e. Patient-Specific Skin Fibroblasts)
Reprogramming mRNA
Direct Conversion Reprogramming Differentiation
TRA-1-60 Oct-4
Cryopreservation
Characterization Left: NSC stained with Nestin (green) and Sox2 (red)
Features:
- Our non-integrating method ensures the absence of a genomic footprint: ideal for drug discovery and cell therapy applications. - Two methods are available: DNA episomal vectors containing the reprogramming factors or synthetic mRNA of each reprogramming factor. - A high reprogramming efficiency is achieved with both the mRNA (0.5%) and the episomal vectors (0.2%). - Both reprogramming vectors and mRNA are removed during the cell cycle. - Safety: both episomal vectors and mRNA are safe to handle. Normal & Patient-Specific Disease Samples & iPSCs Further Characterization Services Available: Now available online! - Karyotyping - Embryoid Body (EB) Formation and Characterization to test iPSC pluripotency Cells / cDNA / Total RNA - Teratoma Formation Analysis Service to test iPSC pluripotency - Neurological Disorders - Oncogenic Disorders iPSC Generation Service (mRNA) ASC-6022 $10,000 - Muscular Disorders iPSC Generation Service (Episomal) ASC-6023 $10,000 - Genetic Disorders - Autoimmune Disorders and more.
iPS Cells
Differentiation
Other Options Oligodendrocytes Differentiation
Characterization Cardiomyocytes Hepatocytes
Dopaminergic Neurons
Skeletal Muscle
Satisfaction Guaranteed! Contact us with information about your samples for details and pricing
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Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Stem Cell Research Products iPS Cell Generation Kit (Retrovirus)
ESC/iPSC Characterization Kit
ASC’s iPS Cell Generation Kit contains an optimized ratio of retroviral reprogramming vectors that has been modified for the consistent generation of iPS cells*. A user-friendly protocol along with our excellent customer service will make the task of iPS cell generation worry-free.
Kit Includes: 1. Easy-to-use primary and secondary antibodies with well-established pluripotency markers to
Features: High Reprogramming Efficiency User-Friendly Protocol Every batch of retrovirus is tested for its ability to generate iPSC colonies Kit Includes: 1. A ready-to-use cocktail of retroviral reprogramming factors (Oct4, Sox2, Klf4 and LMyc) The concentration of each vector has been optimized to obtain a high reprogramming efficiency.
2. Retro-GFP This GFP-containing vector is used for the determination of the optimal condition to maximize the transduction efficiency.
3. Polybrene To enhance efficiency of transduction. 4. Complete medium Just mix the medium supplements and Basal hiPS cell medium included in the kit. 5. Feeder cells CF-1 MEFs are provided for candidate clone culturing
Complete iPS Cell Generation Kit ASK-3011 (up to 4 reactions)
Virus
DAPI
$1,200
Best Selling
confirm pluripotency of human/mouse ES/iPS cell lines. Primary antibodies Human: OCT4, SOX2, SSEA4, TRA-1-60, TRA-1-81 Mouse: OCT4, SOX2, SSEA1
2. Fixation, Permeabilization, Blocking, and Mounting Reagents 3. DNA counterstain 4. Alkaline Phosphatase activity assay
Features: Sophisticated Marker Selection: Each kit includes an optimal panel of pluripotency markers. Marker selection reference table: SSEA-4
TRA-1-60
Human ES cell
Oct-4 +
Sox-2 +
+
+
TRA-1-81 +
Human Neural Stem Cell
-
+
-
-
-
Human Mesenchymal Stem Cell
-
-
+
-
-
Oral SCC9 β6 Cells
-
-
-
+
Human Amniotic Mesenchymal Stem Cell
+
-
-
-
AP +
+ -
+
High Sensitivity: Partially differentiated hiPSC clones can be detected.
Merge
GFP Hoechst staining
Merged
Convenient: The ready-to-use kit contains all the buffers and solutions needed to perform immunocytochemical and enzyme activity analyses.
hOCT4
Mouse ESC/iPSC Characterization Kit (20 reactions) Human ESC/iPSC Characterization Kit (20 reactions)
Top row: BJ cells (Human Fibroblasts) were transduced for 24h with Retro-GFP (MOI: 0.2).
hSOX2
Individual retrovirus is also available: Human iPSC VSVg Retrovirus Cocktail High Titer GFP VSVg Retrovirus
Anti-SSEA-4 staining
ASK-3005 ASK-3006
$200 $290
ES/iPS Cell Pluripotency RT-PCR Kit Easy-to-use RT-PCR kit to confirm the pluripotency of human/mouse ES/iPS cell lines with well established pluripotency markers.
0.4 mL ASR-2005 $1,000 0.1 mL ASR-2006 $230
Mouse ESC/iPSC Pluripotency RT-PCR Kit (20 reactions) ASK-6001 Human ESC/iPSC Pluripotency RT-PCR Kit (20 reactions) ASK-6101
$50 $50
References: *K. Takahashi et al., Cell. 131, 861-72 (2007) 12
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Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
MEF Feeder Cells (Mouse Embryonic Fibroblasts)
Neural Stem Cell Generation Kit Looking for an alternative to expensive protein neural-induction reagents? Tired of the heterogeneous nature of embryoid body formation and the subsequent protracted length of neuronal differentiation? ASC’s Neural Stem Cell Generation Kit is your answer!
Mouse Embryonic Fibroblast (MEF) cells are used as feeder layers for ES/iPS cell cultures. We have a broad selection of high-quality MEF products including: CF-1 Neomycin-resistant DR4 (resistant to neomycin, hygromycin, puromycin, and 6- thioguanine)
Features:
- Fast, efficient, and user-friendly - Special formula: Increases the efficiency of rosette formation by 10-fold over standard neuron and/or induction media - Generation of proliferating Neural Stem Cells (NSCs) that can give rise to large quantities of neuronal and glial cells - Suitable for the generation of various neural cells (glial cells, astrocytes, dopaminergic neurons, motor neurons)
ASK-4002 Neural Stem Cell Generation & Characterization Kit ASK-4003 Neural Stem Cell Generation Kit ASK-4001 Neuro-Sure™ Induction Medium Size: 100 mL ASK-4101 Neuro-Sure™ Culture Medium Size: 100 mL
$700 $500 $150 Inquire
Neuro-Sure™ Medium: Number of Rosette Clusters / Well Standard NIM 3
Neuro-Sure™ (ASM-4001) 30 a) A single Rosette cluster is obeserved using standard neural induction medium. b) Multiple rosette clusters are observed using Neuro-Sure™ Medium.
- Average viability >95% - Tested for long term culture of either mouse or human ES/iPSC lines - Manufactured at our US facility - Rapid inventory turnover ensures fresh stocks are shipped within 2-3 days - Our MEFs have been used by more than 200 clients in both academic and industry labs. DR4, P2, untreated DR4, P3, irradiated
ASF-1001 ASF-1013 ASF-1015
1 x 10^6 cells $180 4 x 10^6 cells $120 2 x 10^6 cells $70
CF-1, P2, untreated CF-1, P3, irradiated
ASF-1201 ASF-1213 ASF-1215
1 x 10^6 cells 4 x 10^6 cells 2 x 10^6 cells
$90 $45 $35
CF-1, P3, mitomycin-C treated ASF-1223 ASF-1225
4 x 10^6 cells 2 x 10^6 cells
$45 $35
Neo-resistant, P2, untreated Neo-resistant, P3, irradiated
1 x 10^6 cells 4 x 10^6 cells 2 x 10^6 cells
$90 $45 $35
ASF-1101 ASF-1113 ASF-1115
SNL SNL 76/7 feeders, STO cell line, P12, untreated SNL 76/7 feeders, STO cell line, P14, irradiated
ASF-1305 ASF-1317
2 x 10^6 cells 5 x 10^6 cells
CF-1
$90 $45
SNL 76/71, 2,3, derived from the mouse fibroblast line STO, is transformed with both the neomycin and LIF genes and has been shown to support the growth of both ES and iPS cell lines. References: 1. Okita, K; Ichisaka, T; Yamanaka, S. Nature 448:313–317. (2007) 2.Takahashi K, Okita K, Nakagawa M, Yamanaka S. Nat Protoc. 2:3081-9. (2007) 3. Takahashi K, Narita M, Yokura M, Ichisaka T, Yamanaka S. PLoS One 4(12):e8067 (2009)
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Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
ESC/iPSC Culture Media
Germline Transmission Tested ESC-Sure™ Fetal Bovine Serum (FBS)
MEF Conditioned Medium ASC’s Conditioned Medium is harvested from cultured CF-1 MEFs. Our stringent quality control and functional control assays ensure the pluripotency of your ES/iPSC lines will be maintened in this feeder-free culture system. ASM-5008 100 ml $120
ESC-Sure™ Fetal Bovine Serum (FBS) is the only serum product on the market that is tested for germline transmission of mouse ES cells.
Serum- and Feeder-Free Medium (SFFM) ESC-Sure™ SFFM is an optimized medium formulated for the culture of human ES/iPSC lines requiring serumfree and feeder-free culture conditioins. ASM-5010 100 ml $99 Mouse ESC Culture Supplement ESC-Sure™ mESC Mate is a basal medium supplement that contains all the factors required by mESC lines. ASM-5011 100 ml $200
Germline transmission tested! Ready-to-use (no heat inactivation needed) Toxin-free
Each lot of ESC-Sure™ FBS is put through a series of quality control tests in addition to a series of functional tests. These tests ensure that each lot of ESC-Sure™ supports the normal morphology, karyotype, pluripotency, and germline transmission of mouse ESCs. Moreover, the series of quality control tests are conducted over several freeze/thaw cycles, therefore ensuring that every lot of ESC-Sure™ purchased is of the highest quality. ESC-Sure™ Fetal Bovine serum, Germline transmission tested ASM-5006 ESC-Sure™ Fetal Bovine serum, Germline transmission tested ASM-5007
Mouse ESC Complete Medium ESC-Sure™ mESC Complete Medium is a ready-to-use medium that contains all the reagents required for mouse ESC and iPSC cultures. ASM-5022 500ml $180
Mouse ES cells cultured in ESC-Sure™ FBS
Neural Stem Cell Differentiation Medium Neuro-Sure Differentiation Medium is a specially formulated medium designed to increase the efficiency of the differentiation of ESCs or iPSCs into neural epithelia. The increase in differentiation efficiency is 10-fold higher than those resulting from standard neural induction media formulas. ASM-4011 100ml $120
ASG-2003-1 ASG-2005-1 ASG-2006-1 ASG-2011-1 ASG-2014-1 ASG-2015-1
DKK-1 (Human Cells) 25 ug Frizzled-5 Fc Fusion (Human Cells) 50 ug Frizzled-8 Fc Fusion (Human Cells) 50 ug SFRP5 (E.Coli) 50 ug Wnt-3a (Human Cells) 5 ug Wnt-5a (Human Cells) 5 ug
Notch Pathway ASI-0030-1 ASG-2009-1
DAPT JAG-1 (Synthesized)
Hedgehog Pathway ASG-2012-1
Sonic Hedgehog (Human cells)
$240 $210 $210 $330 $280 $380
TGF-beta Family Pathway
ASG-2001-1 ASG-2002-1 ASG-1012-1 ASG-2010-1 ASG-2013-1
Others 10 mg 100 ug
5 ug
Request samples! We provide free samples of bFGF, Wnt-3a, TGF-beta1, Noggin, and BMP-4. Contact us at info@appliedstemcell.com. Promo # ASCRP08
$180 $110
$260
ASG-0003 ASG-1010-1 ASG-2004-1 ASG-1014-1 ASG-1004-3 ASG-1006-3 ASG-2007-1 ASG-2008-1 ASG-1005-2 ASG-1007-2 ASG-1009-2
Activin A (Human Cells) BMP-4 (Human Cells) BMP-7 (Human Cells) Noggin (Human Cells) TGF-beta1 (Human Cells)
100 ml 500 ml
Mouse derived from injected ES cells (agouti)
$120 $420
Mouse ES cells cultured in ESC-Sure™ FBS maintain normal morphology and pluripotency. ES cells cultured in our FBS are competent for tetraploid complementation.
Germline transmission test by tetraploid complementation
Stem Cell Growth / Differentiation Factors Wnt Pathway
Mycoplasma-free Virus-free Competitively priced
Mouse ES Cell Lines 5 ug 5 ug 10 ug 50 ug 5 ug
$180 $195 $213 $215 $199
GermLine* C57BL/6-EZ GermLine* C57BL/129SvJ GermLine* 129-EZ GermLine* mESC BALB/c EZ mESC C57BL/6-ALBINO
ASE-9007 ASE-9005 ASE-9006 ASE-9008 ASE-9009
2.5 x 10^6 cells 1 x 10^6 cells 2.5 x 10^6 cells 2 .5 x 10^6 cells 1 x 10^6 cells
$900 $795 $795 $600 $795
*Germline transmission guaranteed
bFGF (E.Coli) 50 ug beta-NGF (Human Cells) 10 ug EGF (E .Coli) 50 ug EPO (Human Cells) 10 ug GM-CSF (Human Cells) 10 ug IFN-alpha 2A (Human Cells) 10 ug IGF-I Long R3 (E. Coli) 100 ug Insulin (E. Coli) 1000 ug IL-2 (Human Cells 10 ug M-CSF (Human Cells) 10 ug TNF-alpha (Human Cells) 10 ug
$95 $104 $85 $240 $213 $142 $120 $110 $109 $207 $87
Human iPS Cell Lines Human iPS Cell Lines generated by retrovirus vectors or episomal expression. Human iPS Cells (Retro) Human iPS Cells (Episomal)
ASE-9101 ASE-9102
2x10^5 cells/vial 2x10^5 cells/vial
$850 $1200
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Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
MEF Feeder Cells
ASM-5008 ESC-Sure™ Conditioned Medium for hESC/iPSC 100 mL $120
ASK-3006
Human ESC/iPSC Characterization Kit 1 kit (20 rxn)
ASM-5010 Serum-/Feeder-free Medium for hESC/iPSC
$99
ASB-0001
Alkaline Phosphatase (AP) Test Solution 1.5 mL
$200
ASB-0101
Cell Fixation Solution
100 mL
ASCAP-0171 Wnt/b-Catenin Regulated cDNA Plate Array (h) 4 rxns
$409
$30
ASCAP-0181 Stem Cell Marker cDNA Plate Array (h) 3 rxns
$409
15 mL
$20
ASCAP-1171 Wnt/b-Catenin Regulated cDNA Plate Array (m) 4 rxns
$409 $469
$579
ASF-1001 DR4 MEF, P2, untreated
1 vial of 1x10^6 cells $180
ASF-1002 DR4 MEF, P2, untreated
3 vials of 1x10^6 cells $490
ASM-5011 ESC-Sure™ mESC Mate
ASF-1013 DR4 MEF, P3, irradiated
1 vial of 4x10^6 cells $120
ASM-5012 ESC-Sure™ Embryoid Body Formation Medium
100 mL
$40
ASB-0102
Permeabilization Solution 15 mL
$20
ASCSC-0001 Single Stem Cell Gene Expression Assay Kit 12 rxns
ASF-1014 DR4 MEF, P3, irradiated
5 vials of 4x10^6 cells $550
ASM-5013 ESC-Sure™ Embryoid Body Formation Medium
500 mL $170
ASB-0103
Blocking Solution
15 mL
$30
ASCSK-0001 Nuclear Extraction Kit
ASF-1015 DR4 MEF, P3, irradiated
1 vial of 2x10^6 cells $70
ASM-5016 ESC-Sure™ ES Grade FBS 100 mL
$95
ASB-0104
DNA Staining Solution
5 mL
$20
ASK-6001
mESC/iPSC Pluripotency RT-PCR Kit 1 kit (20 rxns)
$50
ASF-1016 DR4 MEF, P3, irradiated
5 vials of 2x10^6 cells $325
ASM-5017 ESC-Sure™ ES Grade FBS 500 mL
$320
ASB-0105
Anti-Quench Mounting Solution
$20
ASK-6101
hESC/iPSC Pluripotency RT-PCR Kit 1 kit (20 rxns)
$50
100 mL
ASF-1101 Neo-resistant MEF, P2, untreated
1 vial of 1x10^6 cells $90
ASM-5021 mESC Complete Medium
100 mL
$60
ASF-1102 Neo-resistant MEF, P2, untreated
3 vials of 1x10^6 cells $255
ASM-5022 mESC Complete Medium
500 mL
$180
ASF-1113 Neo-resistant MEF, P3, irradiated
1 vial of 4x10^6 cells $45
Cell Culture Plates
ASF-1114 Neo-resistant MEF, P3, irradiated
5 vials of 4x10^6 cells $215
AM-ART201 NAMEX™ HSC Expansion Kit (6-well) 1 kit
ASF-1115 Neo-resistant MEF, P3, irradiated
1 vial of 2x10^6 cells $35
AM-ART202 NAMEX™ HSC Expansion Kit (24-well)1 kit
ASF-1116 Neo-resistant MEF, P3, irradiated
8 vials of 2x10^6 cells $256
5 mL
Reprogramming and Differentiation Human iPSC VSVg Retrovirus Cocktail 0.4 mL
ASR-2006
High Titer GFP VSVg Retrovirus
$1349
ASK-3011
iPS Cell Generation Kit
$1399
ASK-4002 ASK-4003
$1000
$85
1 kit (4 reactions)
$1200
ASG-2004-2 Recombinant Human EGF
250 ug
$180
NSC Generation & Characterization Kit 1 kit (5 rxn)
$700
ASG-2004-3 Recombinant Human EGF
1000 ug
$290
NSC Generation Kit
$500
ASG-2007-1 Recombinant Human IGF-I Long R3 100 ug
$120
ASG-2008-1 Recombinant Human Insulin
1000 ug
$110
ASG-2011-1
50 ug
$330
1 kit (5 reactions)
ASE-9006 GermLine mESC 129-EZ
2.5x10^6 cells
$795
ASKHZ-0002 G4 Dendritic Cell Generation Kit 500 mL
$1299
1 vial of 4x10^6 cells $45
ASE-9007 GermLine mESC C57BL/6-EZ
2.5x10^6 cells
$900
TARGATT™ Series
ASF-1214 CF-1 MEF, P3, irradiated
5 vials of 4x10^6 cells $215
ASE-9008 GermLine mESC BALB/c EZ
2.5x10^6 cells
$600
AST-0001 TARGATT™ H11 Fresh Embryos (FVB) 25-35 embryos
ASF-1215 CF-1 MEF, P3, irradiated
1 vial of 2x10^6 cells $35
ASE-9009 mESC C57BL/6-ALBINO
1x10^6 cells
$795
ASF-1216 CF-1 MEF, P3, irradiated
8 vials of 2x10^6 cells $256
ASE-9101 iPS Cell Line (Retrovirus)
2x10^5 cells
$850
ASF-1217 CF-1 MEF, P3, irradiated
1 vial of 1x10^6 cells $22
ASE-9102 iPS Cell Line (Episomal)
2x10^5 cells
$1200
3 vials of 1x10^6 cells $255
ASF-1213 CF-1 MEF, P3, irradiated
$95
50 ug
$299
ASF-1202 CF-1 MEF, P2, untreated
50 ug
ASG-2004-1 Recombinant Human EGF
ASKHZ-0001 G4 Dendritic Cell Generation Kit 100 mL
ASE-9005 GermLine mESC C57BL6/129SvJ
Recombinant Human bFGF
$230
$795
1 vial of 1x10^6 cells $90
ASG-0003
0.1 mL
1x10^6 cells
ASF-1201 CF-1 MEF, P2, untreated
$99
Growth Factors
ASR-2005
Mouse ES Cell Lines / Human iPS Cells
1 kit (200 rxns)
Recombinant Human SFRP5
ASG-2001-1 Recombinant Human/Mouse Activin A
5 ug
$180
Inquire
ASG-2001-2 Recombinant Human/Mouse Activin A
25 ug
$750
AST-0002 TARGATT™ H11 Frozen Embryos (FVB) 45-50 embryos
Inquire
ASG-2002-1 Recombinant Human BMP-4
5 ug
$195
AST-0011 TARGATT™ H11 Fresh Embryos (B6)
25-35 embryos
Inquire
ASG-2002-2 Recombinant Human BMP-4
25 ug
$960
AST-0012 TARGATT™ H11 Frozen Embryos (B6) 45-50 embryos
Inquire
ASG-2003-1 Recombinant Human DKK-1
25 ug
$240
ASG-2003-2 Recombinant Human DKK-1
100 ug
$980
ASG-2005-1 Recombinant Frizzled-5 Fc Fusion
50 ug
$210
ASF-1223 CF-1 MEF, P3, mytomycin-C treated 1 vial of 4x10^6 cells $45
Antibodies and Characterization Kit
ASF-1224 CF-1 MEF, P3, mytomycin-C treated 5 vials of 4x10^6 cells $215
ASA-0001 Goat-α-mouse, 590 nm excitation
2 mL
$20
AST-0004 TARGATT™ Rosa 26 Frozen Embryos (FVB)45-50 embryos Inquire
ASF-1225 CF-1 MEF, P3, mytomycin-C treated 1 vial of 2x10^6 cells $35
ASA-0003 Goat-α-rabbit, 590 nm excitation
2 mL
$20
AST-1001
TARGATT™ Transgenic Kit (5 microinjections)
$1450
ASF-1226 CF-1 MEF, P3, mytomycin-C treated 8 vials of 2x10^6 cells $256
ASA-0004 Goat-α-mouse, 488 nm excitation
2 mL
$20
AST-1002
TARGATT™ Transgenic Kit (2 microinjections)
$750
ASF-1305 SNL 76/7 cell line, P12, untreated
1 vial of 2x10^6 cells $90
ASA-0006 Goat-α-rabbit, 488 nm excitation
2 mL
$20
AST-2001
TARGATT™ H11 Mouse Tail Lysis PCR Kit 100 rxn
$189
ASF-1317 SNL 76/7 cell line, P14, irradiated
1 vial of 5x10^6 cells $45
ASA-0110 α-Oct4 (h/m), monoclonal
$80
AST-2002
TARGATT™ Rosa26 Mouse Tail Lysis PCR Kit 100 rxn $189
ASF-1318 SNL 76/7 cell line, P14, irradiated
5 vials of 5x10^6 cells $215
ASA-0120 α-Sox2 (h/m), monoclonal
800 uL
$80
ASA-0135 α-SSEA-1 (m), monoclonal 800 uL $150
AST-0003 TARGATT™ Rosa26 Fresh Embryos (FVB) 25-35 embryos Inquire
ASG-2005-2
Recombinant Frizzled-5 Fc Fusion 250 ug
$960
ASG-2006-1 Recombinant Frizzled-8 Fc Fusion
50 ug
$210
ASG-2006-2 Recombinant Frizzled-8 Fc Fusion
250 ug
$960
ASG-2010-1 Recombinant Human Noggin
50 ug
$215
AST-3001 to AST-3017: TARGATT™ plasmids (2 ug). Available upon request.
ASG-2010-2
250 ug
$920
$80
Assays and Arrays
ASG-2012-1 Recombinant Human Sonic Hedgehog
ASA-0150 α-SSEA-4 (m), monoclonal 800 uL
$80
ASCAP-0007 Stem Cell miRNA Array
$180
ASA-0160 α-TRA-1-60 (h), monoclonal 800 uL
$80
ASCMA-0129 Stem Cell-Specific miRNA Plate Assay Kit I
12 rxns
$449
ASM-5002 ESC-Sure™ Human DMEM/F12 10 bottles of 400 mL (Set) $210
ASA-0170 α-TRA-1-81 (h), monoclonal 800 uL
$80
ASCMA-0130 Stem Cell-Specific miRNA Plate Assay Kit II
12 rxns
ASM-5003 Mouse ES Cell Freezing Medium
50 mL
$65
ASA-1011 Anti-WNT-3a (h/m), polyclonal
100 ug
$260
ASCMA-0102 Stem Cell-Specific miRNA Plate Assay Kit III
ASM-5004 Human ES Cell Freezing Medium
50 mL
$68
ASA-1012 Anti-WNT-5a (h/m), polyclonal
100 ug
$260
$80
Media / Serum ASM-4001 Neuro-Sure™ Medium
100 mL
ASM-5001 ESC-Sure™ Mouse DMEM 10 bottles of 450 mL (Set)
800 uL
ASM-5006 ESC-Sure™ FBS, germline-transmission tested 100 mL
$120
ASA-4010 Anti-Nestin (h), monoclonal 800 uL
ASM-5007 ESC-Sure™ FBS, germline-transmission tested 500 mL
$420
ASK-3005 Mouse ESC/iPSC Characterization Kit 1 kit (20 reactions) $399
$260 $199
ASG-2014-1 Recombinant Human WNT-3a
5 ug
$280
$449
ASG-2015-1 Recombinant Human WNT-5a
5 ug
$380
12 rxns
$449
Small Molecules
ASCMA-0131 Stem Cell-Specific miRNA Plate Assay Kit IV 12 rxns
$449
ASCMA-1003 Stem Cell-Associated miRNA Plate Assay Kit 2 rxns
$409
ASCFA-1003 Stem Cell TF Activation Profiling Plate Array I 6 rxns
$599
$519
Recombinant Human TGF-beta 1
5 ug
5 ug
3 reactions
ASG-2013-1
Recombinant Human Noggin
ASG-2009-1 JAG-1 protein active peptide fragment 100 ug
$110
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Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com
Applied StemCell
Toll Free Phone: 1-866-497-4180
www.appliedstemcell.com