Enzyme-Linked ImmunoSorbent Assay See more with KPL ELISA products — Get higher signal and lower background.
ELISA See more with KPL ELISA Products Sometimes it is difficult to distinguish your protein of interest from background. However, with KPL Protein DetectorTM ELISA products you get strong, clean signal every time. KPL’s ELISA products offer consistent, high quality results and enable sensitive immunodetection of protein in a microwell format. Choose from a broad line of sensitive substrates, high quality kits and stand-alone reagents for detecting and quantifying specific antigens in a complex protein mixture. Combine these products with KPL’s extensive line of secondary antibodies to develop reproducible and sensitive ELISAs. • • • •
Substrates for colorimetric or chemiluminescent detection Secondary antibodies unlabeled or labeled with either horseradish peroxidase (HRP) or alkaline phosphatase (AP) Support reagents – blocking solutions, coating buffers, wash solutions, conjugate stabilizers, stop solutions Universal ELISA kits formatted with HRP- or AP-labeled secondary antibodies
The foundation of KPL’s Protein Detector ELISA offering is a broad line of chemiluminescent and chromogenic substrates for use with HRP and AP conjugates that provide varied levels of sensitivity. For highly sensitive colorimetric peroxidase detection, choose SureBlue Reserve™ Microwell Substrate with long shelf life and convenient 1-component format. New FirePhos™ Microwell Substrate expands our selection of sensitive substrates for phosphatase detection. LumiGLO Reserve™ Chemiluminescent Substrate offers the ultimate in sensitivity via chemiluminescent reaction with HRP. No matter which you choose, KPL’s ELISA substrates are all tested to assure lot-to-lot consistency, reproducibility and long-term stability. An extensive line of high-quality secondary antibody conjugates and labeled streptavidin are also available. Each is affinity purified to assure excellent signal-to-noise results in ELISAs. KPL support reagents are optimized for use in microwell ELISA and tested to verify that they meet strict specifications. They include conjugate stabilizers, diluents, blocking and wash solutions, all formulated to ensure accurate and reproducible results. Protein Detector Kits provide a convenient starting point for the development of ELISA protocols. Each kit contains enzyme-labeled secondary antibody, corresponding substrate, as well as unique diluent/blocking and wash solutions.
KPL ELISA products help you to optimize your assay and SEE MORE!
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Praying Mantis
See more with KPL ELISA products — Get higher signal and lower background.
ELISA ELISA Substrates Substrates are the cornerstones for developing ELISAs with high signal and low background. KPL, one of the first companies to produce stable, liquid substrates, offers a variety of standard and proprietary HRP and AP substrates. See Table 1 for a listing of chromogenic and chemiluminescent substrates.
stability, and consistency, making it the ideal substrate for use in most ELISA applications. Superior Sensitivity with SureBlue Reserve
• •
Chromogenic Peroxidase Substrates
• •
Choose from among three TMB (3,3’,5,5’tetramethylbenzidine) products and two ABTS (2,2”-azino-di-(3-ethylbenzthiazoline -6-sulfonate) formulations. All provide guaranteed performance and reliability. Excellent Signal-to-Noise with SureBlueTM
• • •
High sensitivity Consistent lot-to-lot performance minimizes assay re-optimization 2-year stability
KPL’s SureBlue TMB Microwell Substrate is a 1-component formulation. It develops a deep blue soluble product when reacted with HRP-labeled conjugates. SureBlue TMB provides excellent signal-to-noise,
Highest sensitivity: 50% more sensitive than SureBlue TMB (Figure 1) Consistent lot-to-lot performance minimizes assay re-optimization 2-year stability Ready-to-use 1-component formulation
SureBlue Reserve TMB takes ELISA to a new performance level. It offers maximum sensitivity with exceptionally low background enabling detection of low levels of target protein. SureBlue Reserve TMB provides the same great benefits as KPL’s original SureBlue TMB: low lot-to-lot variation, 2-year stability, ready-to-use 1-component solution, and excellent linearity. TMB Microwell Substrate in a Highly Stable 2-Component Formulation
• •
Longest shelf life due to the 2-component system Higher sensitivity than ABTS
TMB Microwell 2-Component Substrate offers excellent stability and sensitivity. It is the most economical choice when ready-to-use convenience is not required. Like other TMB substrates, it rapidly develops a deep blue color that is measurable at 650 nm. Broad Dynamic Range with ABTS Microwell Substrates
•
• •
Excellent linearity; provides the widest working range of any chromogenic HRP substrate Extremely stable Ideal for kinetic assays
KPL’s ABTS Microwell Substrates offer performance, low background, and a wider linear range than other chromogenic substrates. Supplied either as a ready-to-use formulation or as an extremely stable 2-component product, ABTS substrates develop an intense blue-green color in the presence of peroxidase-labeled conjugates. ABTS is recommended when low background is required and moderate sensitivity is acceptable.
Table 1. KPL Protein Detector ELISA Substrates SureBlue Reserve SureBlue TMB 2-C ABTS 1-C ABTS 2-C BluePhos FirePhos pNPP LumiGLO Reserve LumiGLO PhosphaGLO PhosphaGLO Reserve
Sensitivity ••• •• •• • • •• •• • ••••• •••• •••• •••••
Stability ••• ••• •••• •••• ••••• •• •• ••••• •• •• ••• •••
C = Component
Convenience ••••• ••••• ••• ••••• ••• ••• ••• • ••• ••• ••••• ••••• • = Lowest
Enzyme HRP HRP HRP HRP HRP AP AP AP HRP HRP AP AP
Detection Chromo Chromo Chromo Chromo Chromo Chromo Chromo Chromo Chemi Chemi Chemi Chemi
••••• = Highest
Determination Kinetic or Endpoint Kinetic or Endpoint Kinetic or Endpoint Kinetic or Endpoint Kinetic or Endpoint Kinetic or Endpoint Kinetic or Endpoint Kinetic or Endpoint Endpoint Endpoint Kinetic or Endpoint Kinetic or Endpoint
Comparison of SureBlue and SureBlue Reserve
Comparison of AP Colorimetric Substrates for ELISA
1.0 0.5 0.0 0
0.5
BluePhos FirePhos pNPP
20
SureBlue Reserve SureBlue
1.5
Signal: Background
O.D. at 450 nm
2.0
1
1.5
15 10 5
2 0
HRP Concentration (ng/mL)
For economical chromogenic phosphatase detection that provides sensitivity and convenience, KPL offers three substrates that produce a variety of colors, new FirePhos, BluePhos® and pNPP Substrates. Excellent Overall Performance
pNPP Phosphatase Substrate contains para-nitrophenylphosphate. KPL’s formulation provides one of the most sensitive pNPP substrates available. Both kinetic and endpoint determinations can be performed. It is available in tablet form with a separate buffer solution for shelf stability. Figure 3. A Choice of Colors for Phosphatase Detection
•
Ideal for applications where sensitivity greater than pNPP is required • Stable for 24 months • Convenient and quick: mix and use • Wider dynamic range than pNPP • Lower background than pNPP
• •
Wide linear range Excellent stability
800
600
1000
LumiGLO and LumiGLO Reserve substrates are luminol-based. In the presence of hydrogen peroxide, they are converted to an excited intermediate dianion which emits light on return to its ground state. Both LumiGLO Reserve and LumiGLO substrates offer rapid and sensitive detection. The working range of primary antibody that yields an acceptable signal-tonoise ratio is much wider with these substrates than with chromogenic substrates. Both substrates are supplied as 2-component liquids. PhosphaGLOTM and PhosphaGLO
RedPhos
Economy and Performance with pNPP
400
Figure 2. Human IgG was serially diluted in an ELISA plate and detected with goat ani-human IgG labeled with alkaline phosphatase. KPL’s substrates, FirePhos, BluePhos and pNPP were added to the plate. The reactions were stopped with APstop Solution and read at the recommended wavelength for each substrate. The results indicate that FirePhos and BluePhos produce a significantly higher signal-to-noise relative to pNPP.
with FirePhosTM and BluePhos®
FirePhos and BluePhos Microwell Substrates are based on a soluble, proprietary form of 5-bromo-4-chloro-3indolyl phosphate (BCIP) and produce intense colors (Figure 3). Both substrates offer greater sensitivity and linearity of signal than pNPP (Figure 2) and can be used for both kinetic and endpoint ELISAs.
200
Antigen Concentration (ng/mL)
Figure 1. Dilutions of HRP conjugate were added to microplate wells and the conjugate in each well reacted with either SureBlue or SureBlue Reserve. The reactions were stopped with TMB Stop Solution, and optical density readings were taken at 450 nm. Data points represent the average of triplicate samples at each HRP concentration. The data demonstrate greater sensitivity of SureBlue Reserve at low concentrations of HRP.
Chromogenic Phosphatase Substrates
0
BluePhos
pNPP
ReserveTM Chemiluminescent Phosphatase Substrates
Chemiluminescent Substrates for Peroxidase and Phosphatase
• Femtogram sensitivity with PhosphaGLO Reserve • One-component, ready-to-use • Stable at 4o C for two years.
LumiGLO® and LumiGLO Reserve Chemiluminescent Peroxidase
PhosphaGLO Reserve and PhosphaGLO
Substrates
substrates provide great sensitivity and
• Unsurpassed sensitivity with LumiGLO Reserve • Excellent stability • Fast ramp-up of signal • Stable for 24 hours after mixing • Light emission sustained for 1-2 hours
unmatched convenience. No special blockers are required to achieve low background.
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ELISA See More...
ELISA Antibody Conjugates Choose from a wide selection of secondary antibodies to immunoglobulins from over 20 animal species labeled with horseradish peroxidase or alkaline phosphatase, the most widely used enzymes for ELISA signal generation. HRP has been shown to be more sensitive than AP primarily due to its faster catalytic rate. However, HRP reactions may be self-limiting due to substrate inhibition of enzyme. In contrast, AP exhibits a slower catalytic rate, but is not self-limiting. Reaction rates remain linear over long periods of time; therefore, sensitivity can be improved by allowing the reaction to proceed for a longer time. Whatever enzyme conjugate you choose, all antibody conjugates are of the same exceptionally high quality you have come to expect from KPL. As always, you get results with high signal-to-noise.
Universal ELISA Kits KPL’s Protein Detector ELISA Kits are designed to be an excellent and convenient starting point for development of ELISA protocols. Each kit contains all the reagents necessary to establish an ELISA procedure except for primary antibodies. The kits also include a comprehensive manual to facilitate assay development.
HRP ELISA Kits
KPL’s Protein Detector HRP ELISA Kits contain goat anti-mouse IgG and anti-rabbit IgG, or anti-human IgG HRP conjugates along with ABTS substrate, blocking, coating, substrate, stop and wash solutions. AP ELISA Kits
Included in the Protein Detector AP ELISA kits are goat anti-mouse IgG, anti-rabbit IgG, or anti-human IgG AP conjugates as well as BluePhos substrate, blocking, coating, stop and wash solutions.
ELISA Support Reagents
Try KPL’s ELISA Kits and Reagents KPL has extensive experience in antibody purification and the development of stable liquid substrates. This long-standing expertise is the foundation of our line of Protein Detector ELISA systems and ensures reliable product performance from lot-to-lot. Try our kits and reagents and get the results you’re looking for: a strong, clean signal without background, every time. Your eyes aren’t playing tricks on you. You’ve found the finest line of ELISA systems and reagents at KPL.
KPL’s support reagents are optimized for use in microwell ELISA and tested to verify that they meet strict specifications. They include conjugate stabilizers, diluents, blocking and wash solutions, and stop reagents. Blocking and wash solutions have been designed to reduce background and to ensure a clear signal. KPL stabilizers provide convenient storage solutions for your antibody-enzyme conjugates and ensure their performance. Spend your time developing your assay instead of worrying about making reagents. Choose time-tested, stable support reagents from KPL. LumiGLO and BluePhos are registered trademarks and LumiGLO Reserve, PhosphaGLO, PhosphaGLO Reserve, SureBlue Reserve, SureBlue, FirePhos, ReserveAP, BlueSTOP and Protein Detector are trademarks of KPL, Inc.
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BacTrace® Anti-E.coli O157:H7 Antibody labeled with DyLight 488 (green) directly detects E.coli O157:H7. BacTrace Anti-Salmonella (CSA-1) Antibody labeled with DyLight 549 (red) directly detects Salmonella.
Antibodies to Food Pathogens and Infectious Disease Agents BacTrace® Anti-Bacteria Antibodies provide a powerful set of tools for the detection of food pathogens and other infectious disease agents affecting humans and animals. They are highly characterized affinity purified antibodies that offer excellent specificity and sensitivity in high performance immunoassays such as ELISA, Western blotting, lateral flow and microscopic analysis. Antibodies are offered unlabeled, or labeled with peroxidase, phosphatase, fluorescein, biotin or DyLightTM 488.
BacTrace antibodies offer a range of benefits: • Sensitivity: BacTrace polyclonal antibodies are more sensitive than many commercially available monoclonal antibodies. • Specificity: recognize both cell surface and intracellular antigens of bacterial cells. • Reproducibility: high correlation between antibody lots verified by extensive in-house testing. • Versatility: ideal for dual antibody capture and detection assays; widely used in a variety of applications in research and industrial settings.
Power Your Immunoassays
Unique Process Yields Well-defined Antibodies KPL's premium affinity purification process, developed and refined over thirty years, permits the selection of highly sensitive polyclonal antibodies with various specificity characteristics. ISO 9001:2008-certified quality procedures are carried out at more than six different stages of the production cycle, and material that does not meet our high standards for potency and cross-reactivity is rejected.
Broad Spectrum and Serotype-Specific Detection
BacTrace antibodies are polyclonal and represent a mix of different antibodies recognizing multiple epitopes. The aggregate reactivity is higher than that of many monoclonal antibodies that recognize a single epitope, resulting in higher sensitivity compared to competitors. See Figures 1 and 2 below. Sensitivity of BacTrace Anti-E.coli 0157:H7 vs. Competitor Anti-E.coli Antibody
KPL's unique immunization and purification protocols enable the production of both highly specific (Figure 3, Anti-E.coli O157:H7) and broadly reactive (Figure 4, AntiSalmonella Common Structural Antigen (CSA-1) antibodies). Select the antibody that best meets your assay needs. Specificity of BacTrace Anti-E. coli O157:H7 Antibody Figure 3.
O.D.
Sensitivity
Absorbance (450 nm)
Competitor Anti-E.coli O157:H7 KPL Anti-E.coli O157:H7 Competitor Anti-E.coli Monoclonal
Panel Members
Figure 1. Anti-E. coli primary antibodies were all coated at the same concentration. Goat antibodies were detected with a biotin-labeled mouse anti-goat IgG1 monoclonal antibody and the mouse antibody was detected with a biotin-labeled polyclonal goat anti-mouse IgG antibody at equal concentrations followed by a HRP-streptavidin reporter conjugate.
1.0 2.0
Competitor KPL BacTrace
O.D.
1.5
Figure 4.
O.D.O.D. (405nm)
Sensitivity of BacTrace Anti-Legionella Species Antibody vs. Competitor
Broad Specificity of BacTrace Anti-Salmonella Antibody (CSA-1)
0.5 0 0.01
0.1
1.0
Dilution Factor
Figure 2. Colorimetric assay demonstrating the superior sensitivity of BacTrace Anti-Legionella species Antibody in detecting heat-killed L. pneumophila as compared to a leading competitor.
Power Your Immunoassays
Figures 3 and 4. A colorimetric ELISA was performed showing reactivity of BacTrace Anti-E. coli O157:H7 Antibody (top) and Anti-Salmonella Antibody (bottom). BacTrace Anti-E. coli O157:H7 Antibody is highly selective for the O157:H7 serotype and reacts minimally with other potentially cross-reacting serotypes. BacTrace Anti-Salmonella Antibody demonstrates broad reactivity to Salmonella serotypes including those significant in human disease. Anti-Salmonella antibodies show greatest specificity to Salmonella groups B, C, D and E
Antibodies to Food Pathogens
Reproducibility After meeting rigorous standards throughout the production process, antiserum from multiple animals is pooled, minimizing natural serum variability. This results in more standardized antibody lots and increased consistency. Reproducibility of BacTrace Anti-E.coli O157:H7 Antibody Figure 5. 2.50
O.D.O.D.
2.00
Lot A Lot B Lot C
1.50 1.00 0.50
E.
co li E. O1 co 57: li H7 O 15 ( + 7 ) E. :H con co 7 A tro li T l E. O1 CC lot co :H7 35 030 li E. K1 AT 150 988 co E. li co E. 2 A CC (FF E. O1 li O co TC 11 ) co 28a 12 li A C 77 li , 5 T 2 5 E. O1 128 a A CC 540 co 11 b:H TC 25 4 E. li O :H8 A C 92 co 12 A TC 12 2 E. li O 4:N TCC C 1 808 C Ed it col 26: M B 28 wa rob i O K6 AT AA 10 En rd act 29: 0 A CC -17 te sie er NM TC 43 9 ro lla fre A C 89 ba t a u T 1 3 ct rd nd CC 27 Kl er a ii 9 eb si H clo H7 AT 43 5 e M lla afn ac AT CC 892 or p ia ae C 6 ga ne a A C 87 Sa Pro ne um lve TC 23 9 lm vid lla on i A C 66 on en mo ia TC 13 3 Se ell cia rga e A C 047 rra a ty S ni TC 299 tia ph tua i A C 26 m im rt T 27 Sh ar ur ii A CC 73 i c iu T 2 6 St S gel esc m CC 583 ap h l a en A T 3 0 hy ige fle s CC 36 lo lla xn AT 1 72 co s e C 4 Ye cc on ri A C 02 rs us nei TC 147 8 in au A C 5 ia r T 1 6 r u eu C C 20 ck s 2 er AT 11 5 i A C 06 TC C 6 0 5 C 3 29 8 47 3
0.00
Figure 5. KPL tested three lots of BacTrace Anti-E.coli antibody against a series of ATCC organisms in ELISA assays. The three lots yield similar reactivity patterns and demonstrate high correlation lot-to-lot.
E. coli O157:H7 E. coli O157:H7, Molecular Grade Salmonella CSA-1 Salmonella CSA-1, Molecular Grade Listeria species Campylobacter species Shigella species Vibrio species Vibrio cholerae
Antibodies to Infectious Disease Agents Helicobacter pylori Borrelia burgdorferi Borellia species Legionella species Yersinia species
Bacterial Controls E. coli O157:H7 Salmonella typhimurium Listeria genus specific Campylobacter species Helicobacter pylori Borrelia burgdorferi Renibacterium salmoninarum Shigella species Vibrio species Legionella pneumophila Yersinia species
See back of brochure for product listing.
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Ordering Information Description
Unlabeled
HRP
Goat anti-Campylobacter species
01-92-93 1.0 mg
04-92-93 0.1 mg
Goat anti-E. coli O157:H7
01-95-90 1.0 mg
04-95-90 0.1 mg
Goat anti-E. coli O157:H7, Molecular Grade
01-95-90-MG 1.0 mg
Biotin
DyLight 488
FITC
AP
Reserve AP
Positive Controls
02-92-93 0.5 mg
05-92-93 0.1 mg
50-92-93 1.0 mL
16-95-90 0.5 mg
02-95-90 0.5 mg
05-95-90 0.1 mg
50-95-90 1.0 mL
04-90-90 0.1 mg
16-90-90 0.5 mg
02-90-90 0.5 mg
04-91-99 0.1 mg
16-91-99 0.5 mg
02-91-99 0.5 mg
04-90-01 0.1 mg
16-90-01 0.5 mg
072-03-90-01 0.5 mg
50-90-01 1.0 mL
04-90-02 0.1 mg
16-90-02 0.5 mg
072-03-90-02 0.5 mg
50-90-02 1.0 mL
Antibodies to Food Pathogens
Goat anti-Listeria species
01-90-90 1.0 mg
Goat anti-Listeria species, High Sensitivity
01-90-95 1.0 mg
Goat anti-Salmonella Common Structural Antigen (CSA-1)
01-91-99 1.0 mg
Goat anti-Salmonella (CSA-1), Molecular Grade
01-91-99-MG 1.0 mg
Rabbit anti-Shigella species
01-90-01 1.0 mg
Rabbit anti-Vibrio cholerae
01-90-50 0.5 mg
Rabbit anti-Vibrio species
01-90-02 1.0 mg
051-90-90 0.1 mg
05-91-99 0.1 mg
50-90-90 1.0 mL
50-74-01 1.0 mL
Antibodies to Infectious Disease Agents Goat anti-Borrelia burgdorferi
01-97-91 1.0 mg
04-97-91 0.1 mg
02-97-91 0.5 mg
051-97-91 0.1 mg
50-97-91 1.0 mL
Goat anti-Borrelia species
01-97-92 1.0 mg
04-97-92 0.1 mg
02-97-92 0.5 mg
Goat anti-Helicobacter pylori
01-93-94 1.0 mg
04-93-94 0.1 mg
02-93-94 0.5 mg
Rabbit anti-Legionella species
01-90-03 1.0 mg
04-90-03 0.1 mg
16-90-03 0.5 mg
072-03-90-03 0.5 mg
50-90-03 1.0 mL
Rabbit anti-Yersinia species
01-90-04 1.0 mg
04-90-04 0.1 mg
16-90-04 0.5 mg
072-03-90-04 0.5 mg
50-90-04 1.0 mL
01-96-91 1.0 mg
04-96-91 0.1 mg
50-93-94 1.0 mL
Other Anti-Bacteria Antibodies Goat anti-Renibacterium salmoninarum
02-96-91 0.5 mg
05-96-91 0.1 mg
50-96-91 1.0 mL
All BacTrace antibodies are made in goat unless otherwise noted.
BacTrace antibodies have stood the test of time. As one of the first producers of commercial antibodies to bacteria, KPL has been at the forefront of bacterial immunodetection for over two decades. KPL's BacTrace antibodies have served as key tools for life scientists and diagnostic kit manufacturers in food pathology and infectious disease. Today we are dedicated to advancing the field of bacterial detection through a growing line of antibodies and immunoassay controls. Whether conducting basic research or developing commercial kits, KPL is capable of meeting your needs.
To order or for more information on KPL’s line of immunoassay products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com/bactrace.
Gaithersburg, MD Phone: 800.638.3167/301.948.7755 Fax: 301.948.0169 www.kpl.com
ISO 9001:2008 Registered
ML366-03
Power Your Immunoassays
Multiplex fluorescent detection of E. coli O157:H7 (purple), E. coli O26:H11 (green) and E. coli 0111:H8 (red).
Non-O157 STEC Antibodies E. coli serotypes that produce Shiga toxin (Stx) are responsible for numerous outbreaks traced to the consumption of contaminated foods and beverages. Although E. coli O157:H7 is responsible for the majority of human illnesses attributed to E. coli, there are additional Stxproducing E. coli (e.g., E. coli O26, O111, O45, O121, O103, O145) that can also cause hemorrhagic colitis and postdiarrheal hemolytic uremic syndrome (D+HUS). The Centers for Disease Control and Prevention (CDC) has estimated that 85 percent of E. coli O157:H7 infections are foodborne in origin. Foods that have been identified as sources of contamination include ground beef, lettuce, spinach, and water. BacTrace Anti-Bacterial antibodies that detect STEC strains provide a powerful set of tools to identify these specific food pathogens.
KPL’s BacTrace STEC antibodies offer the following benefits: High specificity, minimal cross-reactivity Affinity purified polyclonal antibody yields higher sensitivity Standardized antibody lots provide consistent performance Optimized for ELISA, Western and Immunohistology applications
Power Your Immunoassays
Sandwich ELISA Testing Cross Reactivity of Antibodies These plots depict OD values from a TMB detected HRP CaptureELISA where a 1:100 dilution (~107 CFU/mL) of a heat- killed positive control from various STEC serogroups and S. Typhimurium was used as the analyte. Unlabeled anti-E. coli O157:H7, O26 and O111 antibodies (KPL catalog #s: 01-95-90, 01-95-92, and
Antibodies to Food Pathogens E. coli O26:H11 E.coli O111:H8 E. coli O157:H7 E. coli O157:H7, Molecular Grade Salmonella CSA-1 Salmonella CSA-1, Molecular Grade Listeria species Listeria species, High Sensitivity Campylobacter species Shigella species Vibrio species Vibrio cholerae
01-95-91, respectively) were used as the capture antibodies and HRP labeled anti-E. coli O111 conjugate (KPL catalog # 04-95-91) was used as the reporter. All other assay components were KPL catalog products, following manufacturer’s protocol.
Ordering Information Description
Goat anti-E.coli O26:H11
Unlabeled
HRP
Biotin
DyLight 488
FITC
AP
Positive Controls
01-95-92 1.0 mg
04-95-92 0.1 mg
16-95-92 0.5 mg
072-03-95-92 0.5 mg
50-95-92 1.0 mL
04-95-91 0.1 mg
16-95-91 0.5 mg
072-03-95-91 0.5 mg
50-95-91 1.0 mL
04-95-90 0.1 mg
16-95-90 0.5 mg
Antibodies to Infectious Disease Agents Helicobacter pylori Borrelia burgdorferi Borellia species Legionella species Yersinia species
Bacterial Controls E. coli O26:H11 E.coli O111:H8 E. coli O157:H7 Salmonella Typhimurium Listeria genus specific Campylobacter species Helicobacter pylori Borrelia burgdorferi Renibacterium salmoninarum Shigella species Vibrio species Legionella pneumophila Yersinia species
011-95-92 0.1 mg Goat anti-E-coli O111:H8
01-95-91 1.0 mg 011-95-91 0.1 mg
Goat anti-E. coli O157:H7
01-95-90 1.0 mg
Goat anti-E. coli O157:H7, Molecular Grade
01-95-90-MG 1.0 mg
02-95-90 0.5 mg
05-95-90 0.1 mg
50-95-90 1.0 mL Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com
To order or for more information on KPL’s full line of immunoassay products, contact us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com.
ML432-02 For research use only. © 2011 KPL, Inc. All rights reserved. Dylight is a trademark of Thermo Fisher Scientific and its subsidiaries.
ISO 9001:2008 Registered
Power Your Immunoassays
HRP ELISA Color Substrates Reliable performance and a range of options The success of an ELISA (enzyme-linked immunosorbent assay) depends on a number of factors, including the performance characteristics of the substrate. KPL’s line of peroxidase (HRP) color substrates provides a range of options to match individual perform-
sulfonate) peroxidase substrates offer reliable performance and low background in ELISA. They turn an intense blue-green in the presence of peroxidase that is measureable between 405 and 415 nm.
Choose the substrate that suits your needs:
ance requirements, including high signal and sensitivity, low background, stability and convenience. As the most preferred peroxidase substrate, TMB (3,3’,5,5’ tetramethylbenzidine) provides high sensitivity and low background in microwell immunoassays. KPL offers a choice of three TMB substrates, starting with SureBlueTM (1-Component), ideal for most ELISA applications. Our TMB Peroxidase
• SureBlue ReserveTM TMB Substrate SureBlue Reserve provides maximum sensitivity while maintaining low background, high consistency and 24-month stability in a convenient 1-component formulation.
• SureBlueTM TMB Microwell Substrate SureBlue TMB offers high sensitivity, consistency and stability in a convenient 1-component solution.
Substrate (2-Component) provides equivalent sensitivity at an economical price. At the high end of sen-
• TMB Peroxidase Substrate
sitivity, SureBlue ReserveTM (1-Component ) is the
TMB Peroxidase Substrate offers equivalent performance to SureBlue in an easy mix-and-use, 2-component formulation.
substrate of choice when ultimate performance is required. All three substrates are recommended for both kinetic and endpoint ELISA. Upon reaction
• ABTS Peroxidase Substrate
with peroxidase, they turn blue and are read at 650
ABTS substrates are supplied as stable 1- and 2-component formulations. They are recommended when low background is required and moderate sensitivity is acceptable.
nm. KPL’s ABTS (2,2’-azino-di-(3-ethylbenzthiazoline-6-
Power Your Immunoassays
TMB Stop Solutions Comparison of KPL’s HRP Microwell Substrates • TMB BlueSTOPTM - formulated to stop color development in microwell assays and to maintain the blue color of TMB. Unlike traditional acid-based TMB stopping solutions, the O.D. value of TMB does not change upon addition of TMB BlueSTOP. The stopped substrate is stable for an extended period for reading at 650 nm. A single 650 nm filter can be used pre- and post-stopping. As the graph above demonstrates, our TMB substrates (SureBlue Reserve, SureBlue and TMB) produce high O.D. values, with SureBlue Reserve exhibiting the highest sensitivity. SureBlue and TMB demonstrate lower O.D. values and are comparable in performance. ABTS 1- and 2-component yield lower O.D. values than TMB while exhibiting similar performance. In summary, KPL’s colorimetric substrates provide a range of sensitivity options to meet a variety of needs.
Ordering Information Catalog# 52-00-01 52-00-02 52-00-03 53-00-01 53-00-02 53-00-03 50-76-00 50-76-03 50-66-18 50-66-00 50-66-06 50-62-00 50-62-01
Description SureBlue TMB Microwell Substrate (1-Component) SureBlue TMB Microwell Substrate (1-Comp) SureBlue TMB Microwell Substrate (1-Comp) SureBlue Reserve TMB Microwell Substrate (1-Comp) SureBlue Reserve TMB Microwell Substrate (1-Comp) SureBlue Reserve TMB Microwell Substrate (1-Comp) TMB Microwell Substrate (2-Comp) TMB Microwell Substrate (2-Comp) ABTS Microwell Substrate (1-Comp) ABTS Microwell Substrate (1-Comp) ABTS Microwell Substrate (1-Comp) ABTS Substrate System (2-Comp) ABTS Substrate System (2-Comp)
50-85-30 50-85-41 50-85-05 50-85-06 50-85-01
Stop Solutions TMB BlueSTOP Solution TMB BlueSTOP Solution TMB Stop Solution TMB Stop Solution ABTS Stop Solution
Size 100 mL 400 mL 1L 100 mL 400 mL 1L 600 mL 2700 mL 100 mL 600 mL 1L 600 mL 2700 mL
400 mL 1L 400 mL 1L 200 mL
• TMB Stop Solution - changes the blue color of TMB substrate to yellow and increases absorbance values 2 - 3 times. Stopped TMB is read at a wavelength of 450 nm.
• ABTS Stop Solution - specifically designed for use with KPL’s ABTS Peroxidase Substrates to stop color development in micro-well assays. ABTS retains the green color after stopping.
Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com
ISO 9001:2008 Registered
To order or for more information on KPL’s line of protein detection products, contact us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com. SureBlue, SureBlue Reserve and TMB BlueSTOP are trademarks of KPL. ABTS is a registered trademark of Roche Biochemicals. ©2008 KPL, Inc. All rights reserved. ML352-03
Power Your Immunoassays
SureBlue Reserve™
Take your analysis to a new level of sensitivity
SureBlue Reserve™ TMB ELISA Substrate KPL has expanded its recognized line of ready-to-use TMB ELISA substrates to provide a range of sensitivity options. Our latest addition, SureBlue Reserve, enables you to easily detect your target analyte by providing maximum sensitivity and exceptionally low background. With detection levels of less than one nanogram per milliliter of HRP along with excellent linearity ⎯ SureBlue Reserve provides a superior signal-to-noise ratio, quick results and extremely low lot-to-lot variation. Excellent Signal-to-Noise Ratio
Highly Consistent Lot-to-lot variation is less than 10% ⎯ making the results highly reproducible. SureBlue Reserve eliminates the need for assay re-optimization every time a new lot is used. Long Shelf Life
SureBlue Reserve is highly stable and guaranteed to perform for at least 24 months when stored properly for added economy and convenience. Convenience
SureBlue Reserve is the substrate of choice when highest sensitivity is desired. In low end protein detection, SureBlue Reserve provides at least 50% greater sensitivity than KPL’s SureBlue TMB Substrate as measured by change in O.D. at 450 nm. It detects less than 0.4 ng/ml of HRP with exceptionally low background, less than 0.03 O.D. at 650 nm.
SureBlue Reserve is a one-component, ready-to-use, substrate. Environmentally Safe SureBlue Reserve is water-based and contains no DMF or DMSO.
Power Your Immunoassays
SureBlue Reserve ™
SureBlue Reserve vs. SureBlueTM
Related Products TMB Stop Solution Catalog No. 50-85-05 Size: 400 mL
SureBlue Reserve is the substrate of choice when highest sensitivity is desired with chromogenic substrates. In low end protein detection, SureBlue Reserve provides at least 50% greater sensitivity than SureBlue as measured by the change in optical density units at 450 nm. The data also reveals that SureBlue Reserve's limit of detection is approximately two-fold that of SureBlue.
SureBlue TMB Microwell Substrate (1-comp) Catalog No. 52-00-01 Size: 100 mL
Long Term Stability of SureBlue Reserve
HRPStabilizer Catalog No. 54-15-01 Size: 200 mL
10% BSA Diluent/Blocking Solution Concentrate Catalog No. 50-61-00 Size: 200 mL
Substrate activity and background levels were measured for SureBlue Reserve stored at 4°C over a 28 month period. Activity was measured by reacting the substrate with 2.5 ug, 5 ug, 7.5 ug, or 10 ug of HRP conjugate and measuring optical density (O.D.) at 650 nm. Percent of original activity at 1 month is represented above on the primary axis. Background was measured by O.D. of unreacted substrate at 650 nm and is represented on the secondary axis. Results indicate that SureBlue Reserve is extremely stable over a 28 month period with no significant loss of activity or increase in background when stored at 2-8°C. The data also reveals that any slight change in substrate activity will be more apparent with lowering amounts of HRP.
Coating Solution Concentrate Catalog No. 50-84-00 Size: 50 mL
Wash Solution Concentrate Catalog No. 50-63-00 Size: 800 mL
Ordering Information Catalog#
Description
Size
53-00-01
SureBlue Reserve TMB Microwell Substrate (1-Comp)
100 mL
53-00-02
SureBlue Reserve TMB Microwell Substrate (1-Comp)
4 x 100 mL
53-00-03
SureBlue Reserve TMB Microwell Substrate (1-Comp)
1000 mL
SureBlue Reserve can be provided in a wide variety of custom and bulk packaging options. Contact your KPL sales representative for more details.
Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169
To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.
www.kpl.com
ISO 9001:2008 Registered
SureBlue and SureBlue Reserve are trademarks of KPL, Inc. ML280-06 Power Your Immunoassays
SureBlue™ SureBlueTM TMB 1-Component Microwell Peroxidase Substrate Not all TMB substrates are equal - only one comes with over 20 years of experience. SureBlue TMB is the product of our longstanding expertise in the development of stable liquid substrates. As a single, readyto-use reagent, it offers not only convenience but reliable performance in ELISA applications. SureBlue contains 3,3',5,5' - tetramethylbenzidine in an acidic buffer and develops a deep blue soluble product when reacted with horseradish peroxidase labeled conjugates in microwell plates or tubes. KPL's SureBlue offers the following benefits: Lot-to-lot consistency KPL’s commitment to the strictest quality control standards is realized in the consistency of SureBlue. Lot-to-lot variations do
not exceed 15%. SureBlue provides consistent performance everytime with little need for assay re-optimization between lots.
Stability / Long Shelf-life SureBlue is a highly stable formulation. SureBlue exhibits high activity and low background for at least 24 months from date of manufacture. KPL's maximum acceptable background is 0.03 O.D. at 650 nm.
Flexibility When the SureBlue reaction is stopped by acidification, it remains stopped for 60 minutes. Therefore, there is no need to read the plate immediately. SureBlue provides the flexibility to schedule plate measurements at your convenience.
Power Your Immunoassays
Long Term Stability Substrate activity and background levels were measured for two lots of SureBlue stored at 2-8°C over a 24 month period. Activity was measured by reacting the substrate with serial dilutions of HRP conjugate from 10ng/mL to 0.2 ng/mL. After development, the substrate was stopped and the O.D. measured at 450 nm and compared to the original activity as measured at one month. The substrate’s activity over 24 months compared to its original activity is represented on the primary axis. Background was measured by O.D. of unreacted substrate at Figure 1. 650 nm and is represented on the secondary axis. Results indicate that SureBlue is extremely stable over a 24 month period with no significant loss of activity or increase in background when stored at 2-8°C. (Figure 1). Lot-to-Lot Consistency
Stable Stopped Reaction
Our experience allows us to produce the same high-performance substrate lot after lot.
Once SureBlue is stopped, it stays stopped. This feature provides flexibility in plate measurement of manual assays, and makes it the ideal choice for automation.
For each of nine lots of SureBlue, triplicate samples of HRP anti-human IgG were tested at concentrations ranging from 20-400 pg/well. The reaction was stopped with acid, and the average optical density of the triplicate samples was reported as shown below. Figure 2.
Lot No.
Conclusion: The variation in activity among each lot of KPL’s SureBlue was less than 15%.
Average optical density of stopped triplicate samples (500 pg/well HRP anti-human IgG) were taken at various time points for SureBlue and competitor TMB (see Figure 3). The optical density of each test sample was normalized to its time zero reading. Figure 3.
SureBlue™ Related Products TMB Stop Solution Catalog No. 50-85-05 Size: 400 mL
HRPStabilizer Catalog No. 54-15-01 Size: 200 mL
10% BSA Diluent/Blocking Solution Concentrate Catalog No. 50-61-00 Size: 200 mL
Coating Solution Concentrate Catalog No. 50-84-00 Size: 50 mL
Wash Solution Concentrate Catalog No. 50-63-00 Size: 800 mL
Conclusion: Results demonstrate that stopped SureBlue is the most stable stopped TMB substrates employed in this study.
Ordering Information Catalog No.
Description
52-00-01 52-00-02 52-00-03
SureBlue TMB Microwell Substrate (1-Comp) SureBlue TMB Microwell Substrate (1-Comp) SureBlue TMB Microwell Substrate (1-Comp)
Gaithersburg, MD 20878
Size 100 mL 4 x 100 mL 1000 mL
Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com
ISO 9001:2008 Registered
To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com. SureBlue is a trademark of KPL, Inc. © 2003 KPL, Inc. All rights reserved. ML-242-12 0207
Power Your Immunoassays
Peroxidase Conjugates KPL Peroxidase Conjugates: Time-tested, Sensitive and Reliable Since 1979 KPL has provided quality affinity
streptavidin. They are affinity purified and in
purified antibodies to researchers worldwide.
some cases further adsorbed to minimize cross-
Over the years we have refined our production
reactivity between animal species or shared
process to provide antibodies with high potency
reactivity with other immunoglobulin classes.
and consistent performance in immunoassays.
HRP-labeled F(ab’)2 fragment antibodies are
From the start KPL gives careful consideration
offered for assays requiring extremely low back-
to immunogen preparation, using a highly
ground and absence of F(c)-mediated binding.
purifed formulation to generate antiserum. KPL pools antiserum from multiple animals to
KPL reacts HRP of the highest quality with
reduce natural animal to animal serum variabili-
affinity purified antibodies and streptavidin
ty. During the purification process our ISO
using the periodate method of Nakane and
9001:2008-certified quality procedures impose
Kawaoi. Special features of HRP include:
rigorous standards for potency and cross-reac-
• faster catalytic rate than alkaline phosphatase • generates more product in shorter incubation
tivity. The result is standardized antibody lots with excellent reproducibility. Our extensive line of peroxidase (HRP) conjugates is available across a range of animal species, including human, mouse, rabbit and rat antibodies, as well as other animal species and
times
• provides maximum sensitivity, low nonspecific binding
• ideal for ELISA, Western blotting and immunohistology applications. Power Your Immunoassays
Peroxidase Conjugates Peroxidase Conjugates Ordering Information (Partial listing) Catalog#
Description
Size
KPL offers a range of sensitive substrates for use with HRP conjugates. They provide a choice of intense colors for ELISA, blotting and cell staining applications.
04-10-06
HRP-labeled Goat Anti-Human IgG (H+L)
0.1 mg
04-10-17
HRP-labeled Goat Anti-Human IgA+IgG+IgM (H+L), MSA
0.1 mg
04-10-20
HRP-labeled Goat Anti-Human IgG (Fc)
0.1 mg
074-1002
HRP-labeled Goat Anti-Human IgG (γ)
1.0 mg
074-1003
HRP-labeled Goat Anti-Human IgM (μ)
1.0 mg
074-1004
HRP-labeled Goat Anti-Human IgE (ε)
1.0 mg
074-1006
HRP-labeled Goat Anti-Human IgG (H+L)
1.0 mg
074-1007
HRP-labeled Goat Anti-Human IgA+IgG+IgM (H+L)
1.0 mg
14-10-01
HRP-labeled Goat Anti-Human IgA (α)
0.5 mg
214-1002
HRP-labeled F(ab’)2 Goat Anti-Human IgG (γ)
0.5 mg
214-1003
HRP-labeled F(ab’)2 Goat Anti-Human IgM (μ)
0.5 mg
214-1006
HRP-labeled F(ab’)2 Goat Anti-Human IgG (H+L)
0.5 mg
474-1002
HRP-labeled Goat Anti-Human IgG (γ), Liquid
1.0 mL
474-1003
HRP-labeled Goat Anti-Human IgM (μ), Liquid
1.0 mL
474-1006
HRP-labeled Goat Anti-Human IgG (H+L), Liquid
1.0 mL
•
4 CN Peroxidase Substrate
04-18-06
HRP-labeled Goat Anti-Mouse IgG (H+L), HSA
0.1 mg
•
04-18-15
HRP-labeled Goat Anti-Mouse IgG (H+L), RtSA, HSA
0.1 mg
TMB Membrane Peroxidase Substrate
04-18-18
HRP-labeled Goat Anti-Mouse IgG (H+L), RbSA, HSA
0.1 mg
074-1802
HRP-labeled Goat Anti-Mouse IgG (γ), HSA
1.0 mg
074-1803
HRP-labeled Goat Anti-Mouse IgM (μ), HSA
1.0 mg
074-1806
HRP-labeled Goat Anti-Mouse IgG (H+L), HSA
1.0 mg
074-18-061
HRP-labeled Goat Anti-Mouse IgG (H+L), XSA
1.0 mg
074-1807
HRP-labeled Goat Anti-Mouse IgA+IgG+IgM (H+L), HSA
1.0 mg
074-1809
HRP-labeled Goat Anti-Mouse IgG+IgM (H+L), HSA
1.0 mg
14-18-01
HRP-labeled Goat Anti-Mouse IgA (α), HSA
0.5 mg
214-1802
HRP-labeled F(ab’)2 Goat Anti-Mouse IgG (γ), HSA
0.5 mg
214-1806
HRP-labeled F(ab’)2 Goat Anti-Mouse IgG (H+L), HSA
0.5 mg
474-1802
HRP-labeled Goat Anti-Mouse IgG (γ), HSA, Liquid
1.0 mL
474-1806
HRP-labeled Goat Anti-Mouse IgG (H+L), HSA, Liquid
1.0 mL
074-1506
HRP-labeled Goat Anti-Rabbit IgG (H+L)
1.0 mg
074-15-061
HRP-labeled Goat Anti-Rabbit IgG (H+L), XSA
1.0 mg
074-1516
HRP-labeled Goat Anti-Rabbit IgG (H+L), HSA
1.0 mg
214-1516
HRP-labeled F(ab’)2 Goat Anti-Rabbit IgG (H+L), HSA
0.5 mg
474-1506
HRP-labeled Goat Anti-Rabbit IgG (H+L), Liquid
1.0 mL
474-1516
HRP-labeled Goat Anti-Rabbit IgG (H+L), HSA, Liquid
1.0 mL
14-30-00
HRP-labeled Streptavidin
0.5 mg
Phone: 800.638.3167
474-3000
HRP-labeled Streptavidin, Liquid, Molecular Grade
1.0 mL
Fax: 301.948.0169
Visit our website at www.kpl.com for a complete listing of HRP-labeled antibodies. To order or for more information on KPL’s protein research products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com. For research use only. ©2009 KPL, Inc. All rights reserved. ABTS is a registered trademark of Roche Biochemicals. ML371-02
ELISA •
ABTS® 1- and 2-Component Microwell Peroxidase Substrates
•
SureBlueTM TMB Peroxidase Substrate
•
SureBlue ReserveTM TMB Peroxidase Substrate
•
TMB Peroxidase Substrate
Blotting
•
•
LumiGLO®Chemiluminescent Substrate LumiGLO ReserveTM Chemiluminescent Substrate
Whichever substrate you choose, enjoy the benefits of excellent signalto-noise and reproducibility.
Gaithersburg, MD 20878
www.kpl.com
ISO 9001:2008 Registered
Power Your Immunoassays
AP ELISA Color Substrates See your ELISA results in vivid colors! Prefer vibrant red or brilliant blue? Choose between new FirePhosTM, BluePhos® or pNPP AP Microwell Substrates for sensitive ELISA detection! KPL offers a choice of three sensitive substrates for the detection and quantification of phosphatase (AP) activity in microwell assays. New FirePhosTM AP Microwell Substrate BluePhos® AP Microwell Substrate pNPP Phosphatase Substrate Whichever substrate you choose, enjoy the benefits of excellent signal-to-noise with higher sensitivity and lower background than that of other AP microwell substrates. They produce strong colors that can be read visually. When combined with KPL’s APstopTM Solution, signal development is completely stopped for endpoint determinations. The fast rate of color generation also permits accurate, quantitative measurement in kinetic ELISA.
Extended stability All three AP substrates are stable for one year. Convenience FirePhos and BluePhos are offered as liquid two-component solutions. To prepare mix equal parts and use. Sensitivity FirePhos and BluePhos provide greater sensitivity than pNPP and can detect less than 0.5 pg of phosphatase. (See data on reverse side.) KPL’s pNPP substrate is one of the most sensitive pNPP formulations available, showing enhanced performance over competitive products. The color produced is stable and linear over a wide dynamic range. Reproducibility All KPL AP colorimetric substrates are optimized for consistent results from lot-to-lot, assay-to-assay. Power Your Immunoassays
ReserveAP Conjugates
Phosphatase Substrates for ELISA
TM
Improve your assay performance even further with high potency ReserveAPTM Antibodies KPL’s new ReserveAPTM Alkaline Phosphatase-labeled Conjugates exhibit high potency and consistent performance in immunoasComparison of AP colorimetric substrates for ELISA Human IgG was serially diluted in an ELISA plate and detected with goat anti-human IgG labeled with alkaline phosphatase. KPL’s substrates, FirePhos, BluePhos, pNPP and a competitor pNPP were added to the plate. The reactions were stopped with APstop Solution and read at the recommended wavelength for each substrate. The results indicate that FirePhos and BluePhos produce a significantly higher signal-to-noise relative to pNPP.
result of advances in our conjugation technology and offer higher signal and lower background than our current line of alkaline phos-
Ordering Information Catalog#
says. These conjugates are the
phatase conjugates while meeting
Description
Size
the same standards for stability and reproducibility. They are ideal
FirePhosTM AP Microwell Substrate
for immunoassays that require
50-87-10
FirePhos Microwell Phosphatase Substrate System
50 mL
50-87-20
FirePhos Microwell Phosphatase Substrate System
200 mL
high detection sensitivity in an AP
50-87-30
FirePhos Microwell Phosphatase Substrate System
600 mL
system, including ELISA, Western
50-87-40
FirePhos Microwell Phosphatase Substrate System
2.7 L
blotting and immunohistology.
BluePhos® AP Microwell Substrate 50-88-02
BluePhos Phosphatase Substrate for ELISA
50 mL
50-88-00
BluePhos Phosphatase Substrate for ELISA
600 mL
50-88-01
BluePhos Phosphatase Substrate for ELISA
2700 mL
pNPP Phosphatase Substrates 50-80-00
pNPP Phosphatase Substrate Kit, 100 tabs & DEA Buffer
500 mL
50-80-01
pNPP Substrate, 5 mg tablets
100 tabs
Stop Solutions 50-89-00
Gaithersburg, MD 20878
APstop Solution (10X) (formerly BluePhos Stop Solution)
200 mL
Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com ISO 9001:2008 Registered
To order or for more information, contact us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com. ML347-03 For research use only. BluePhos is a trademark of KPL and is covered by U.S. Patent No. 5,916,746. © 2007 KPL, Inc. All rights reserved.
Power Your Immunoassays
ReserveAP Conjugates TM
TM
Spice up your assay with our red hot high potency ReserveAP Conjugates! KPL’s new ReserveAPTM Alkaline Phosphatase
line of AP conjugates in ELISA and outperform
(AP)-labeled antibody conjugates exhibit high
AP conjugates offered by other manufacturers.
potency and consistent performance in
Higher conjugate dilutability is also observed
immunoassays. These conjugates are the result
without loss of linearity, enabling precious
of advances in our conjugation technology and
antigen or primary antibody conservation.
offer higher signal than our current line of AP
Consistent Performance
conjugates while meeting the same standards for low background, stability and reproducibili-
Reproducible antibody conjugation and consis-
ty. They are intended for demanding
tent performance are verified according to our
immunoassays that require high detection sen-
ISO 9001:2008-registered quality management
sitivity, including ELISA, Western blotting and
system. Lot consistency studies in which three
immunohistology.
lots were studied by ELISA indicated minimal variability.
Higher Potency Excellent Value ReserveAP Conjugates are affinity purified and conjugated to the highest grade of alkaline
ReserveAP Conjugates provide high perform-
phosphatase. In our studies, they generate two-
ance at an economical price.
to-three fold higher values than our current Power Your Immunoassays
AP Substrates Chemiluminescent ELISA: A constant amount of antigen (mouse IgG) was coated onto a microtiter plate. The plate was probed with varying concentrations of goat anti-mouse IgG (H+L) AP conjugate, including KPL’s standard AP conjugate and ReserveAP™ conjugate, followed by KPL’s PhosphaGLO™ Chemiluminescent AP Substrate.
KPL offers a range of sensitive substrates for the detection and quantification of phosphatase (AP) activity. They provide a choice of intense colors for ELISA and blotting applications.
ELISA •
Conclusion: The results indicate that ReserveAP™ conjugate offers superior performance to our standard AP conjugate with a ~3-fold lower amount of conjugate required for detection.
ReserveAPTM Conjugates Ordering Information
•
•
Catalog#
Description
Size
0751-1001
Goat Anti-Human IgA (α)
1.0 mg
0751-1002
Goat Anti-Human IgG (γ)
1.0 mg
0751-1003 0751-1004
Goat Anti-Human IgM (µ) Goat Anti-Human IgE (ε)
0751-1006
FirePhosTM AP Microwell Substrate BluePhos® AP Microwell Substrate pNPP Phosphatase Substrate
Blotting •
1.0 mg 1.0 mg
FirePhos AP Membrane Substrate
•
BCIP/NBT Phosphatase Substrate
Goat Anti-Human IgG (H+L)
1.0 mg
•
PhosphaGLO AP Substrate
0751-1007
Goat Anti-Human IgA+IgG+IgM (H+L)
1.0 mg
•
2151-1002
0.5 mg
4751-1002
F(ab’)2 Anti-Human IgG (γ) Goat Anti-Human IgG (γ) liquid
PhosphaGLOTM Reserve AP Substrate
4751-1003
Goat Anti-Human IgM (µ) liquid
1.0 mg
4751-1006
Goat Anti-Human IgG (H+L) liquid
1.0 mg
0751-1802
Goat Anti-Mouse IgG (γ) HSA
1.0 mg
0751-1803
Goat Anti-Mouse IgM (µ) HSA
1.0 mg
0751-1806
Goat Anti-Mouse IgG (H+L) HSA
1.0 mg
0751-1809
Goat Anti-Mouse IgG +IgM (H+L) HSA
1.0 mg
4751-1802
Goat Anti-Mouse IgG (γ) HSA, liquid
1.0 mg
4751-1806
Goat Anti-Mouse IgG (H+L) HSA, liquid
1.0 mg
151-18-01
Goat Anti-Mouse IgA (α) HSA
0.5 mg
0751-1807
Goat Anti-Mouse IgA+IgG+IgM (H+L) HSA
1.0 mg
0751-1506
Goat Anti-Rabbit IgG (H+L)
1.0 mg
0751-1516
Goat Anti-Rabbit IgG (H+L) HSA
1.0 mg
4751-1506
Goat Anti-Rabbit IgG (H+L), liquid
1.0 mg
4751-1516
Goat Anti-Rabbit IgG (H+L) HSA, liquid
1.0 mg
1.0 mg
HSA=Human Serum Adsorbed
Whichever substrate you choose, enjoy the benefits of excellent signalto-noise with higher sensitivity and lower background than that of other AP substrates. Visit www.kpl.com for more information.
Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com
ISO 9001:2008 Registered
Visit our website at www.kpl.com for a complete listing of ReserveAP conjugates. To order or for more information on KPL’s protein research products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com. ML349-06 For research use only. ©2007 KPL, Inc. All rights reserved.
Power Your Immunoassays
ELISpot ELISpot Protein DetectorTM ELISpot Kits Protein Detector ELISpot kits are designed to provide complete, flexible systems for conducting ELISpot assays with any matched pair of antibodies. For the beginner or the more experienced researcher, these kits are ideal due to their comprehensive reagents and optimized protocol. Convenience Each kit contains all reagents for ELISpot detection except for matched antibody pairs (unlabeled coating and biotinylated antibody). Stable, ready-to-use, one-component substrates and uncoated Multiscreen速 PVDF Plates are included. Other kit components are supplied as concentrates; simply dilute and use. Stability All kit reagents are stable for a minimum of one year, providing reliable signal development and consistently low background.
Visualization Visualizing immunospots has never been easier with the exceptional localization and resolution provided by kit substrates. For sharp localization and fine resolution of bright blue immunospots, choose the HRP kit with TrueBlueTM substrate. For excellent visibility from a dark purple precipitate, choose the AP kit with BCIP/NBT substrate. Flexibility Universal nature of reagents allows kits to be used with any matched pair of antibodies. Since labeled streptavidin is compatible with any biotinylated antibody, the researcher simply provides the antibody pair for detection of different cytokines or other secreted proteins. Additional flexibility is offered through two kit formats. Choose between the HRP kit with TrueBlue or the AP kit with BCIP/NBT. Power Your Immunoassays
Image above. Mouse spleen cells secreting IFN-gamma were detected in ELISpot with either BCIP/NBT (top rows) or TrueBlue Substrate (bottom rows).
ELISpot
Protein Detector HRP and AP ELISpot Kits Protein DetectorTM ELISpot Kits offer pretested reagents and a convenient, optimized protocol for ELISpot protein detection via either horseradish peroxidase (HRP) or alkaline phosphatase (AP) detection systems. With all necessary reagents except the paired antibody set (unlabeled capture antibody and biotinylated detection antibody), this kit allows for minimal time and effort in developing a sensitive and reproducible ELISpot assay. Provided conjugates can detect any biotinylated antibody making these reagents easily interchangeable with any matched antibody pair.
Related Products *TrueBlue Peroxidase Substrate Catalog No. 71-00-64 Size: 50 mL
*BCIP/NBT Substrate (1-Comp) Catalog No. 50-81-18 Size: 100 mL
Kit contents: • Chromogenic Substrate HRP Kit: TrueBlue AP Kit: BCIP/NBT • Labeled Streptavidin HRP Kit: Peroxidase labeled Streptavidin AP Kit: Phosphatase labeled Streptavidin • 5 Multiscreen PVDF Plates, 96-well • Coating Solution Concentrate • BSA Blocking/Diluent Solution Concentrate • Washing Solution Concentrate
HRP-labeled Streptavidin Catalog No. 474-3000 Size: 1.0 mL
AP-labeled Streptavidin Catalog No. 475-3000 Size: 1.0 mL
Comparison of Substrates for ELISpot Color Development
Substrate
Format
Enzyme
Sensitivity
TrueBlue
1-C
HRP
Blue
√√√√
BCIP/NBT
1-C
AP
Dark Purple
√√
AEC
2-C
HRP
Red
√√
C= Component
√=Least sensitive
√√√√=Most Sensitive
*BSA Diluent/Blocking Solution Concentrate Catalog No. 50-61-00 Size: 200 mL
Coating Solution Concentrate Catalog No. 50-84-00 Size: 50 mL
Wash Solution Concentrate Catalog No. 50-63-00 Size: 800 mL
* Other sizes available
Ordering Information Catalog#
Description
Size
40-00-05
Protein Detector HRP ELISpot Kit
5 plates
50-00-05
Protein Detector AP ELISpot Kit
5 plates
Each kit includes substrate (TrueBlue or BCIP/NBT), HRP or AP-labeled Streptavidin, 5 Multiscreen PVDF Plates, 96-well, Coating Solution Concentrate, BSA Blocking/Diluent Solution Concentrate, and Washing Solution Concentrate. Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169
To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.
www.kpl.com
ISO 9001:2008 Registered
TrueBlue is a registered trademark and Protein Detector and Detector are trademarks of KPL, Inc. Multiscreen is a registered trademark of Millipore Corporation. © 2003 KPL, Inc. All rights reserved. ML290-06
Power Your Immunoassays
Immunoassay Buffers Save preparation time with KPL’s new line of quality buffers! Lighten your work load with KPL’s new line of quality
registered quality management system. All buffers are con-
buffers. Offered as convenient liquid concentrates, they
ductivity controlled. KPL offers a variety of buffers designed
eliminate the need for time-consuming buffer preparation.
for use in general immunoassay applications such as Western
KPL buffers are manufactured and carefully controlled for
blotting, gel electrophoresis, ELISA and sample preparation.
quality and consistent performance with our ISO 9001:2008Description
1X Composition
Applications
10X Tris-Glycine Transfer Buffer
25 mM Tris, 192 mM glycine. pH 8.3
10X Tris-Glycine-SDS
25 mM Tris, 192 mM glycine, 0.1% SDS. pH 8.3
For preparing a standard Western blot transfer buffer (Towbin) and as a gel electrophoresis buffer for native Tris-glycine gels without SDS. . Running buffer for sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS-PAGE) of proteins.
10X Phosphate-Buffered Saline (PBS)
10 mM Phosphate,150 mM NaCl. pH 7.4
Phosphate-Buffered Tween 20, (PBST)
10 mM Phosphate, 150 mM NaCl, 0.05% Tween 20. pH 7.4
10X Tris-Buffered Saline (TBS)
50 mM Tris, 150 mM NaCl. pH 7.6
10% Tween 20
10% Tween 20
Wash buffer for general immunoassay applications and sample preparation. Where applicable, Tween 20 is a nonionic detergent that reduces nonspecific binding and protein-protein interaction during wash steps.
Nonionic detergent additive for PBS and Western blotting wash solutions. Reduces nonspecific binding and proteinprotein interactions.
See reverse side.
Power Your Immunoassays
Immunoassay Buffers
KPL Quality Buffers (cont’d) Description
1X Composition
Applications
5% SDS
5% Sodium Dodecyl Sulfate
Detergent surfactant used in preparing proteins for sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS-PAGE) and as an additive to transfer buffers in Western blotting. SDS increases the elution rate of proteins from the gel.
20X SSC
15 mM sodium citrate, 150 mM NaCl in DEPC water. pH 7.0
Used in a variety of molecular biology applications. Facilitates transfer of nucleic acids to membranes.
10X Dulbecco's PBS (D-PBS)
8.5 mM sodium phosphate, 1.5 mM potassium phosphate, 137 mM NaCl. pH 7.4
For sample preparation and as a wash buffer in general immunoassay, tissue, and cell culture applications. Not formulated with magnesium or calcium salts. Where applicable, Tween 20 is a non-ionic detergent that reduces nonspecific binding.
Other Immunoassay Support Reagents Available from KPL ELISA and Western Blotting Applications: • AP and HRP Conjugate Stabilizers
10X Dulbecco's with Tween 20. (D-PBST)
8.5 mM sodium phosphate, 1.5 mM potassium phosphate, 137 mM NaCl, 2.7 mM KCl. 0.05% Tween 20. pH 7.4
10X Tris-Buffered Saline with 0.5% Tween 20 (TBST)
50 mM Tris, 150 mM NaCl, 0.05% Tween 20. pH 7.6
10X Tris-Buffered Saline with 10% Tween 20 (TBST)
50 mM Tris, 150 mM NaCl, 1.0% Tween 20. pH 7.6
• 10% BSA Diluent/Blocking Solution Concentrate • Milk/Diluent Blocking Solution Concentrate • 20X Wash Solution Concentrate
As a general wash buffer in immunoassay applications. Tween 20 is a non-ionic detergent that reduces nonspecific binding. The 10% Tween 20 formulation provides a more stringent wash than standard TBST formulations.
Western Blotting Applications: • Detector Block • Biotin Wash Kit
Take the time and effort out of your buffer preparation. Choose from our line of popular buffers and enjoy the benefits - convenience, reliability and confidence - that KPL’s pretested, quality buffers bring to your research.
ELISA Applications: • Coating Solution Concentrate • Stop Solutions
Ordering Information Catalog#
Description
Size
51-10-01 51-10-02 51-11-01 51-11-02 51-13-01 51-13-02 51-14-01 51-14-02 51-17-01 51-17-02 51-12-01 51-12-02 51-20-01 50-86-05 51-15-01 51-15-02 51-16-01 51-16-02 51-18-01 51-18-02 51-19-01 51-19-02
10X Tris-Glycine Transfer Buffer
1L 5L 1L 5L 1L 5L 200 mL 1L 1L 5L 200 mL 1L 200 mL 1L 1L 5L 200 mL 1L 200 mL 1L 200 mL 1L
10X Tris-Glycine SDS Buffer 10X Phosphate-Buffered Saline (PBS) 10X Phosphate-Buffered Saline with Tween 20 (PBST) 10X Tris-Buffered Saline (TBS) 10% Tween 20 5% SDS 20X SSC 10X Dulbecco's PBS (D-PBS) 10X Dulbecco's PBS with Tween 20 (D-PBST) 10X Tris Buffered Saline with 0.5% Tween 20 (TBST) 10X Tris Buffered Saline with 10% Tween 20 (TBST)
Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com
ISO 9001:2008 Registered
To order or for more information on KPL’s full line of protein detection products, contact us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com. ©2009 KPL, Inc. All rights reserved. ML363-04
Power Your Immunoassays
Ordering Information
Catalog #
Description
Size
Catalog #
Description
Protein Detector ELISA Kits
Phosphatase Chemiluminescent Substrates
Each kit includes an AP- or HRP-labeled Conjugate, Coating Solution, Wash Solution Concentrate, 10% BSA Diluent/Blocking Solution Concentrate, Stop Solution and Substrate.
55-60-03
PhosphaGLO AP Substrate
55-60-04
Same as above
55-60-01
PhosphaGLO Reserve AP Substrate
Phosphatase
55-60-02
Same as above
55-81-10
AP ELISA Kit, Anti-Human
20 plates
55-81-50
AP ELISA Kit, Anti-Rabbit and Anti-Mouse
20 plates
Size
30 mL 100 mL 30 mL 100 mL
Antibody Conjugates
54-62-10
HRP ELISA Kit, Anti-Human
20 plates
All antibodies listed below are produced in goat. For a complete antibody listing, visit www.kpl.com or refer to KPL's Product Catalog.
54-62-18
HRP ELISA Kit, Anti-Mouse
20 plates
Phosphatase
54-62-15
HRP ELISA Kit, Anti-Rabbit
20 plates
Peroxidase
075-1006
ReserveAP Anti-Human IgG (H+L)
Substrates for ELISA
075-1806
ReserveAP Anti-Mouse IgG (H+L), HSA
1.0 mg
Phosphatase Chromogenic Substrates
075-1506
ReserveAP Anti-Rabbit IgG (H+L)
1.0 mg 1.0 mg
50 mL
1.0 mg
50-88-02
BluePhos Microwell Substrate Kit
50-88-00
Same as above
600 mL
074-1006
Anti-Human IgG (H+L)
50-88-01
Same as above
2.7 L
074-1806
Anti-Mouse IgG (H+L), HSA
1.0 mg
50-87-10
FirePhos Microwell Substrate Kit
50 mL
074-1506
Anti-Rabbit IgG (H+L)
1.0 mg 0.5 mg
Peroxidase
50-87-20
Same as above
200 mL
Biotin-Labeled
50-87-30
Same as above
600 mL
16-10-06
Anti-Human IgG (H+L)
50-87-40
Same as above
2.7 L
16-18-06
Anti-Mouse IgG (H+L), HSA
0.5 mg
50-80-00
pNPP Phosphatase Substrate System
500 mL
16-15-06
Anti-Rabbit IgG (H+L)
0.5 mg
50-80-01
pNPP Phosphatase Tablets
100 tablets
Labeled Streptavidin
Peroxidase Chromogenic Substrates
15-30-00
AP-labeled
0.5 mg
52-00-01
SureBlue TMB 1-Component Substrate
100 mL
14-30-00
HRP-labeled
0.5 mg
52-00-02
Same as above
400 mL
52-00-03
Same as above
1L
53-00-01
SureBlue Reserve TMB 1-Component Substrate
100 mL
Blocking Solutions
Same as above
400 mL
50-61-00
10% BSA Diluent/Blocking Solution
200 mL
1L
50-61-10
10% BSA Diluent/Blocking Solution
1L
600 mL
50-82-01
Milk Diluent/Blocking Solution
53-00-02
Assay Support Reagents 50-84-00
Coating Solution Concentrate
50-63-00
Wash Solution Concentrate
50 mL 800 mL
53-00-03
Same as above
50-76-00
TMB 2-Component Substrate Kit
50-76-03
Same as above
50-66-18
ABTS Microwell Substrate 1-Component
55-15-00
APStabilizer
200 mL
100 mL
54-15-01
HRPStabilizer
200 mL
Stop Solutions
2.7 L
50-66-00
Same as above
600 mL
50-66-06
Same as above
1L
50-62-00
ABTS Substrate System 2-Component
50-62-01
Same as above
600 mL 2700 mL
Peroxidase Chemiluminescent Substrates 54-61-00
LumiGLO Chemiluminescent Substrate Kit
240 mL
54-61-01
Same as above
720 mL
54-61-02
Same as above
60 mL
54-71-00
LumiGLO Reserve Chemiluminescent Substrate Kit
54-71-01
ML299-03
Same as above
50-85-01
ABTS Stop Solution
200 mL
50-89-00
APstop Solution
200 mL
50-85-05
TMB Stop Solution
400 mL
50-85-06
TMB Stop Solution
50-85-30
TMB BlueSTOP Solution
400 mL
50-85-41
TMB BlueSTOP Solution
1L
2400 cm2 600 cm2
200 mL
Conjugate Stabilizers
HSA: Human serum adsorbed
To order, call 800.638.3167 or 301.948.7755, fax 301.948.0169, online at www.kpl.com or contact your local KPL Sales Partner. ISO 9001:2008 Registered
1L