Enzyme-Linked ImmunoSorbent Assay. See more with KPL ELISA products Get higher signal and lower background. SEE MORE with KPL!

Transcription

1 Enzyme-Linked ImmunoSorbent Assay See more with KPL ELISA products Get higher signal and lower background.

2 ELISA See more with KPL ELISA Products Sometimes it is difficult to distinguish your protein of interest from background. However, with KPL Protein Detector TM 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!

3 Praying Mantis See more with KPL ELISA products Get higher signal and lower background.

4 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. 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 SureBlue TM 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, stability, and consistency, making it the ideal substrate for use in most ELISA applications. Superior Sensitivity with SureBlue Reserve 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 Sensitivity Stability Convenience Enzyme Detection Determination SureBlue Reserve HRP Chromo Kinetic or Endpoint SureBlue HRP Chromo Kinetic or Endpoint TMB 2-C HRP Chromo Kinetic or Endpoint ABTS 1-C HRP Chromo Kinetic or Endpoint ABTS 2-C HRP Chromo Kinetic or Endpoint BluePhos AP Chromo Kinetic or Endpoint FirePhos AP Chromo Kinetic or Endpoint pnpp AP Chromo Kinetic or Endpoint LumiGLO Reserve HRP Chemi Endpoint LumiGLO HRP Chemi Endpoint PhosphaGLO AP Chemi Kinetic or Endpoint PhosphaGLO Reserve AP Chemi Kinetic or Endpoint C = Component = Lowest = Highest

5 Comparison of SureBlue and SureBlue Reserve Comparison of AP Colorimetric Substrates for ELISA 2.0 O.D. at 450 nm SureBlue Reserve SureBlue Signal: Background BluePhos FirePhos pnpp HRP 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 Antigen Concentration (ng/ml) 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. Chromogenic Phosphatase Substrates 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 with FirePhos TM and BluePhos 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 FirePhos and BluePhos Microwell Substrates are based on a soluble, proprietary form of 5-bromo-4-chloro-3- indolyl 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. Economy and Performance with pnpp Wide linear range Excellent stability 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 RedPhos BluePhos pnpp Chemiluminescent Substrates for Peroxidase and Phosphatase LumiGLO and LumiGLO Reserve Chemiluminescent Peroxidase Substrates 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 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. PhosphaGLO TM and PhosphaGLO Reserve TM Chemiluminescent Phosphatase Substrates Femtogram sensitivity with PhosphaGLO Reserve One-component, ready-to-use Stable at 4 o C for two years. PhosphaGLO Reserve and PhosphaGLO substrates provide great sensitivity and unmatched convenience. No special blockers are required to achieve low background

6 ELISA ELISA Antibody Conjugates See More... 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 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. 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. 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

7 HRP ELISA Kits Protein Detector TM Horseradish Peroxidase (HRP) ELISA Kit Antibody Sandwich (Capture) ELISA Indirect Antibody ELISA Direct ELISA Hybridoma Screening Protein Detector TM ELISA Kit HRP System offers a convenient starting point for the development of microwell procedures for antigen or antibody detection. The kit provides pretested components including HRP-labeled anti-mouse and anti-rabbit or anti-human secondary antibodies along with blocking, coating and wash solutions which substantially reduce the time and effort needed to develop a reproducible assay. This kit also contains ABTS, a peroxidase substrate that provides reliability, convenience and excellent overall performance. A comprehensive manual for developing enzyme immunoassay procedures is also provided. Users only need to provide primary antibody and plates to run an ELISA. Protein Detector ELISA kits offer the following benefits: Excellent Dynamic Range Excellent linearity: ABTS provides the widest working range of any chromogenic HRP substrate. Extremely Stable All kit components are stabilized for ease-ofuse and reliability. Very low background, long shelf life. Convenient Non-hazardous, no special handling or disposal required. Optimized liquid stystem makes your ELISA experiments faster and easier. Ideal for use in a variety of ELISA techniques.

8 HRP ELISA Kits Performance of ABTS Substrate Features of ABTS Substrate Absorbance ABTS unstopped ABTS stopped Substrate Type Kit Components Detection Limit Kinetics Stability of Working Solution Hazards Color Before Stopping ABTS Chromogenic moles of HRP Slow 24 hours None Blue-Green Each kit contains: 1. Coating Solution Concentrate 2. BSA Diluent/Blocking Solution Concentrate 3. Wash Solution Concentrate 4. 50% glyceral Absorbance Before Stopping nm 5. HRP-labeled antibody,.1 mg Stop Reagent Color After Stopping ABTS Stop Solution Blue-Green 6. 2-component ABTS Substrate Conjugate Concentration (ng/ml) Absorbance After Stopping Amplification After Stopping nm None 7. Stop Solution Concentrate 8. Manual ABTS offers a broad working range prior to stopping and after stopping. Ordering Information Storage: Store kit reagents at 2-8 C. Stability: Kit is stable for a minimum of 1 year from date of receipt. Catalog No. Description Size Protein Detector ELISA Kit, HRP System 2000 tests Anti-Human Antibody Protein Detector ELISA Kit, HRP System 2000 tests Anti-Mouse Antibody Protein Detector ELISA Kit, HRP System 2000 tests Anti-Rabbit Antibody Related Products Protein Detector ELISA Kit, AP System 2000 tests Anti-Mouse and Anti-Rabbit Antibodies Protein Detector ELISA Kit, AP System 2000 tests Anti-Human Antibody To order or for more information on KPL s full line of protein and nucleic acid detection products, contact us at / , FAX or visit us at Gaithersburg, MD Phone: Fax: ISO 9001:2000 Registered ML ABTS is a registered trademark of the Roche Group KPL, Inc. All rights reserved.

9 AP ELISA Kits Protein Detector TM Alkaline Phosphatase (AP) ELISA Kits Antibody Sandwich (Capture) ELISA Indirect Antibody ELISA Direct ELISA Hybridoma Screening Protein Detector AP ELISA Kits offer an excellent and convenient starting point for the development of ELISA protocols. These kits provide pretested goat anti-mouse, antirabbit or anti-human AP conjugates as well as blocking, coating and wash solutions. KPL kits substantially reduce the effort needed to develop a sensitive and reproducible assay. All components needed to establish an ELISA except for primary antibodies are provided. AP ELISA kits utilize BluePhos, a phosphatase substrate that provides superior performance over pnpp. A comprehensive manual for developing enzyme immunoassay procedures is also provided. Protein Detector AP ELISA kits offer the following benefits: Highly Sensitive Detect at least 2 ng of protein and less than 0.5 pg of phosphatase in free solution. Extremely Stable Very low background, long shelf life. Excellent Dynamic Range BluePhos offers improved signal-to-noise ratio and linearity over pnpp. Convenient Optimized liquid system makes your ELISA experiments faster and easier. Nonhazardous, no special handling or disposal required. Ideal for use in a variety of ELISA techniques.

10 Performance Comparison of BluePhos and pnpp AP ELISA Kits O.D. Ordering Information 3.0 BluePhos 2.5 pnpp Natural Log (Dilution) Catalog No. Description Size Protein Detector ELISA Kit, AP System 2000 tests Anti-Mouse and Anti-Rabbit Antibodies Protein Detector ELISA Kit, AP System 2000 tests Anti-Human Antibody Related Products Protein Detector ELISA Kit, HRP System 2000 tests Anti-Human Antibody Methodology Mouse IgG was coated onto two different ELISA plates. The starting dilution for the BluePhos plate was 1 mg/ml, while the pnpp plate started at 10 μg/ml. The antigen was diluted 2-fold eight times. Phosphatase-labeled goat antimouse IgG was added to each well at a concentration of 500 ng/ml. The plate was washed, and substrate applied. Based on the background readings at time 0, and the final readings at time 15 minutes, the amount of protein was determined by dividing the dilution concentration by 100 μl (the volume used when coating the well). Therefore, the amount of protein detected by BluePhos (point 7) is 1.56 ng, while the amount detected by pnpp (point 5) is 62.4 ng. Conclusion The graph shows the superiority of BluePhos in both increased sensitivity and in producing a wider dynamic range than pnpp Protein Detector ELISA Kit, HRP System 2000 tests Anti-Mouse Antibody Protein Detector ELISA Kit, HRP System 2000 tests Anti-Rabbit Antibody Each kit contains: Coating Buffer Concentrate, BSA Diluent/Blocking Solution Concentrate, Wash Solution Concentrate, 50% Glycerol Solution, Enzyme-labeled Secondary Antibody, 2-Component Chromogenic Substrate System (Substrate components may vary depending upon kit.), and Stop Solution. To order or for more information on KPL s full line of protein and nucleic acid detection products, contact us at / , FAX or visit us at Gaithersburg, MD Phone: Fax: ISO 9001:2000 Registered BluePhos is a registered trademark and Protein Detector is a trademarks of KPL, Inc KPL, Inc. All rights reserved. ML271-06

11 Immunoassay Buffers Save preparation time with KPL s new line of quality buffers! Lighten your work load with KPL s new line of quality buffers. Offered as convenient liquid concentrates, they eliminate the need for time-consuming buffer preparation. KPL buffers are manufactured and carefully controlled for quality and consistent performance with our ISO 9001:2008- registered quality management system. All buffers are conductivity controlled. KPL offers a variety of buffers designed for use in general immunoassay applications such as Western blotting, gel electrophoresis, ELISA and sample preparation. Description 1X Composition Applications 10X Tris-Glycine 25 mm Tris, 192 mm glycine. For preparing a standard Western blot transfer buffer Transfer Buffer ph 8.3 (Towbin) and as a gel electrophoresis buffer for native Tris-glycine gels without SDS.. 10X Tris-Glycine-SDS 25 mm Tris, 192 mm glycine, Running buffer for sodium dodecyl sulfate polyacrylamide 0.1% SDS. ph 8.3 gel electrophoresis (SDS-PAGE) of proteins. 10X Phosphate-Buffered 10 mm Phosphate,150 mm NaCl. Wash buffer for general immunoassay applications Saline (PBS) ph 7.4 and sample preparation. Where applicable, Tween 20 is a nonionic detergent that reduces nonspecific Phosphate-Buffered 10 mm Phosphate, 150 mm NaCl, binding and protein-protein interaction during wash steps. Tween 20, (PBST) 0.05% Tween 20. ph X Tris-Buffered Saline 50 mm Tris, 150 mm NaCl. (TBS) ph % Tween 20 10% Tween 20 Nonionic detergent additive for PBS and Western blotting wash solutions. Reduces nonspecific binding and proteinprotein interactions. See reverse side.

12 KPL Quality Buffers (cont d) Description 1X Composition Applications 5% SDS 5% Sodium Detergent surfactant used in preparing proteins for Dodecyl Sulfate 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, Used in a variety of molecular biology applications. 150 mm NaCl Facilitates transfer of nucleic acids to membranes. in DEPC water. ph X Dulbecco's 8.5 mm sodium phosphate, For sample preparation and as a wash buffer in general PBS (D-PBS) 1.5 mm potassium phosphate, immunoassay, tissue, and cell culture applications. 137 mm NaCl. ph 7.4 Not formulated with magnesium or calcium salts. Where applicable, Tween 20 is a non-ionic detergent that reduces nonspecific binding. 10X Dulbecco's 8.5 mm sodium phosphate, with Tween 20. (D-PBST) Ordering Information 1.5 mm potassium phosphate, 137 mm NaCl, 2.7 mm KCl. 0.05% Tween 20. ph X Tris-Buffered 50 mm Tris, 150 mm NaCl, As a general wash buffer in immunoassay applications. Saline with 0.5% 0.05% Tween 20. ph 7.6 Tween 20 is a non-ionic detergent that reduces nonspecific Tween 20 (TBST) binding. The 10% Tween 20 formulation provides a more stringent wash than standard TBST formulations. 10X Tris-Buffered 50 mm Tris, 150 mm NaCl, Saline with 10% 1.0% Tween 20. ph 7.6 Tween 20 (TBST) 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. Immunoassay Buffers Other Immunoassay Support Reagents Available from KPL ELISA and Western Blotting Applications: AP and HRP Conjugate Stabilizers 10% BSA Diluent/Blocking Solution Concentrate Milk/Diluent Blocking Solution Concentrate 20X Wash Solution Concentrate Western Blotting Applications: Detector Block Biotin Wash Kit ELISA Applications: Coating Solution Concentrate Stop Solutions Catalog# Description Size X Tris-Glycine Transfer Buffer 1 L L X Tris-Glycine SDS Buffer 1 L L X Phosphate-Buffered Saline (PBS) 1 L L X Phosphate-Buffered Saline with 200 ml Tween 20 (PBST) 1 L X Tris-Buffered Saline (TBS) 1 L L % Tween ml L % SDS 200 ml X SSC 1 L X Dulbecco's PBS (D-PBS) 1 L L X Dulbecco's PBS with Tween 20 (D-PBST) 200 ml L X Tris Buffered Saline with 0.5% Tween ml (TBST) 1 L X Tris Buffered Saline with 10% Tween ml (TBST) 1 L To order or for more information on KPL s full line of protein detection products, contact us at / , fax or visit us at Gaithersburg, MD Phone: Fax: ISO 9001:2008 Registered 2009 KPL, Inc. All rights reserved. ML363-03

13 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 performance 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 SureBlue TM (1-Component), ideal for most ELISA applications. Our TMB Peroxidase Substrate (2-Component) provides equivalent sensitivity at an economical price. At the high end of sensitivity, SureBlue Reserve TM (1-Component ) is the substrate of choice when ultimate performance is required. All three substrates are recommended for both kinetic and endpoint ELISA. Upon reaction with peroxidase, they turn blue and are read at 650 nm. KPL s ABTS (2,2 -azino-di-(3-ethylbenzthiazoline-6- 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: SureBlue Reserve TM TMB Substrate SureBlue Reserve provides maximum sensitivity while maintaining low background, high consistency and 24-month stability in a convenient 1-component formulation. SureBlue TM TMB Microwell Substrate SureBlue TMB offers high sensitivity, consistency and stability in a convenient 1-component solution. TMB Peroxidase Substrate TMB Peroxidase Substrate offers equivalent performance to SureBlue in an easy mix-and-use, 2-component formulation. ABTS Peroxidase Substrate ABTS substrates are supplied as stable 1- and 2-component formulations. They are recommended when low background is required and moderate sensitivity is acceptable.

14 Comparison of KPL s HRP Microwell Substrates TMB Stop Solutions TMB BlueSTOP TM - 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# Description Size SureBlue TMB Microwell Substrate (1-Component) 100 ml SureBlue TMB Microwell Substrate (1-Comp) 400 ml SureBlue TMB Microwell Substrate (1-Comp) 1 L SureBlue Reserve TMB Microwell Substrate (1-Comp) 100 ml SureBlue Reserve TMB Microwell Substrate (1-Comp) 400 ml SureBlue Reserve TMB Microwell Substrate (1-Comp) 1 L TMB Microwell Substrate (2-Comp) 600 ml TMB Microwell Substrate (2-Comp) 2700 ml ABTS Microwell Substrate (1-Comp) 100 ml ABTS Microwell Substrate (1-Comp) 600 ml ABTS Microwell Substrate (1-Comp) 1 L ABTS Substrate System (2-Comp) 600 ml ABTS Substrate System (2-Comp) 2700 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. Stop Solutions TMB BlueSTOP Solution 400 ml TMB BlueSTOP Solution 1 L TMB Stop Solution 400 ml TMB Stop Solution 1 L ABTS Stop Solution 200 ml To order or for more information on KPL s line of protein detection products, contact us at / , fax or visit us at Gaithersburg, MD Phone: Fax: ISO 9001:2000 Registered SureBlue, SureBlue Reserve and TMB BlueSTOP are trademarks of KPL. ABTS is a registered trademark of Roche Biochemicals KPL, Inc. All rights reserved. ML352-02

15 AP ELISA Color Substrates See your ELISA results in vivid colors! Prefer vibrant red or brilliant blue? Choose between new FirePhos TM, 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 FirePhos TM 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 APstop TM 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.

16 Phosphatase Substrates for ELISA ReserveAP TM Conjugates Improve your assay performance even further with high potency ReserveAP TM Antibodies Comparison 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. Ordering Information Catalog# Description Size FirePhos TM AP Microwell Substrate FirePhos Microwell Phosphatase Substrate System 50 ml FirePhos Microwell Phosphatase Substrate System 200 ml FirePhos Microwell Phosphatase Substrate System 600 ml FirePhos Microwell Phosphatase Substrate System 2.7 L KPL s new ReserveAP TM Alkaline Phosphatase-labeled Conjugates exhibit high potency and consistent performance in immunoassays. These conjugates are the result of advances in our conjugation technology and offer higher signal and lower background than our current line of alkaline phosphatase conjugates while meeting the same standards for stability and reproducibility. They are ideal for immunoassays that require high detection sensitivity in an AP system, including ELISA, Western blotting and immunohistology. BluePhos AP Microwell Substrate BluePhos Phosphatase Substrate for ELISA 50 ml BluePhos Phosphatase Substrate for ELISA 600 ml BluePhos Phosphatase Substrate for ELISA 2700 ml pnpp Phosphatase Substrates pnpp Phosphatase Substrate Kit, 100 tabs & DEA Buffer 500 ml pnpp Substrate, 5 mg tablets 100 tabs Stop Solutions APstop Solution (10X) (formerly BluePhos Stop Solution) 200 ml Gaithersburg, MD Phone: Fax: ISO 9001:2000 Registered To order or for more information, contact us at / , fax or visit us at ML For research use only. BluePhos is a trademark of KPL and is covered by U.S. Patent No. 5,916, KPL, Inc. All rights reserved.

17 ReserveAP TM Conjugates Spice up your assay with our red hot high potency ReserveAP TM Conjugates! KPL s new ReserveAP TM Alkaline Phosphatase (AP)-labeled antibody conjugates exhibit high potency and consistent performance in immunoassays. These conjugates are the result of advances in our conjugation technology and offer higher signal than our current line of AP conjugates while meeting the same standards for low background, stability and reproducibility. They are intended for demanding immunoassays that require high detection sensitivity, including ELISA, Western blotting and immunohistology. Higher Potency ReserveAP Conjugates are affinity purified and conjugated to the highest grade of alkaline phosphatase. In our studies, they generate twoto-three fold higher values than our current line of AP conjugates in ELISA and outperform AP conjugates offered by other manufacturers. Higher conjugate dilutability is also observed without loss of linearity, enabling precious antigen or primary antibody conservation. Consistent Performance Reproducible antibody conjugation and consistent performance are verified according to our ISO 9001:2000-registered quality assurance system. Lot consistency studies in which three lots were studied by ELISA indicated minimal variability. Excellent Value ReserveAP Conjugates provide high performance at an economical price.

18 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 FirePhos TM AP Microwell Substrate 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. ReserveAP TM Conjugates Ordering Information Catalog# Description Size Goat Anti-Human IgA (α) 1.0 mg Goat Anti-Human IgG (γ) 1.0 mg Goat Anti-Human IgM (µ) 1.0 mg Goat Anti-Human IgE (ε) 1.0 mg Goat Anti-Human IgG (H+L) 1.0 mg Goat Anti-Human IgA+IgG+IgM (H+L) 1.0 mg F(ab ) 2 Anti-Human IgG (γ) 0.5 mg Goat Anti-Human IgG (γ) liquid 1.0 mg Goat Anti-Human IgM (µ) liquid 1.0 mg Goat Anti-Human IgG (H+L) liquid 1.0 mg Goat Anti-Mouse IgG (γ) HSA 1.0 mg Goat Anti-Mouse IgM (µ) HSA 1.0 mg Goat Anti-Mouse IgG (H+L) HSA 1.0 mg Goat Anti-Mouse IgG +IgM (H+L) HSA 1.0 mg Goat Anti-Mouse IgG (γ) HSA, liquid 1.0 mg Goat Anti-Mouse IgG (H+L) HSA, liquid 1.0 mg Goat Anti-Mouse IgA (α) HSA 0.5 mg Goat Anti-Mouse IgA+IgG+IgM (H+L) HSA 1.0 mg Goat Anti-Rabbit IgG (H+L) 1.0 mg Goat Anti-Rabbit IgG (H+L) HSA 1.0 mg Goat Anti-Rabbit IgG (H+L), liquid 1.0 mg Goat Anti-Rabbit IgG (H+L) HSA, liquid 1.0 mg HSA=Human Serum Adsorbed Visit our website at for a complete listing of ReserveAP conjugates. To order or for more information on KPL s protein research products, contact us at / , FAX or visit us at ML For research use only KPL, Inc. All rights reserved. BluePhos AP Microwell Substrate pnpp Phosphatase Substrate Blotting FirePhos AP Membrane Substrate BCIP/NBT Phosphatase Substrate PhosphaGLO AP Substrate PhosphaGLO TM Reserve AP Substrate Whichever substrate you choose, enjoy the benefits of excellent signalto-noise with higher sensitivity and lower background than that of other AP substrates. Visit for more information. Gaithersburg, MD Phone: Fax: ISO 9001:2000 Registered

19 BacTrace Anti-Bacteria Antibodies BacTrace E.coli O157:H7 Antibody labeled with DyLight 488 (green) directly detects E.coli O157:H7. BacTrace Salmonella (CSA-1) Antibody labeled with DyLight 549 (red) directly detects Salmonella. (DyLight labeling performed on a custom basis.) BacTrace Antibodies to Food Pathogens and Other Infectious Agents KPL produces a unique line of BacTrace antibacteria antibodies for the detection of food pathogens and other infectious agents affecting humans and animals. They are purified to offer excellent specificity and sensitivity in high performance immunoassays such as ELISA, Western blotting, lateral flow and microscopic analysis. Many leading diagnostic kit manufacturers use BacTrace antibodies in their diagnostic kits. See our growing line of BacTrace antibodies: Anti-Salmonella Anti-Salmonella, Molecular Grade Anti-E.coli O157:H7 Anti-E.coli O157:H7, Molecular Grade Anti-Listeria, genus-specific Anti-Campylobacter, genus specific Anti-Helicobacter pylori Anti-Borrelia species Anti-Borrelia burgdorferi Anti-Renibacterium salmoninarum BacTrace antibodies offer a range of benefits: Sensitivity: nanogram detection of bacterial antigens within hours. Specificity: recognize both cell surface and intracellular antigens of bacterial cells. Versatility: ideal for screening assays in the research lab, environmental monitoring and for food and beverage analysis in the industrial setting. Positive controls available for each BacTrace antibody. Anti-Salmonella and E.coli O157:H7, Molecular Grade New molecular grade antibodies to Salmonella and E. coli O157:H7 are recent additions to our BacTrace line. These antibodies are specially processed and QC-tested to ensure that contaminating Salmonella or E. coli DNA is not detectable by real-time PCR or other molecular methods.

20 KPL has a large production capacity and offers flexible services to meet your specific needs. Multi-gram quantities from the same purification lot Labeling options, including peroxidase, phosphatase, fluorescein Customized antibody formulations to tailor specificity and sensitivity Custom conjugation to other reporter molecules (fluorophores, enzymes, etc.) Ordering Information Catalog# Description Size Anti-Salmonella (CSA-1) Common Structural Antigen Unlabeled 1.0 mg MG Unlabeled, molecular grade 1.0 mg Phosphatase-labeled 0.1 mg Peroxidase-labeled 0.1 mg Fluorescein-labeled 0.5 mg Salmonella typhimurium Positive Control Antigen 1.0 ml Anti-E. coli O157:H Unlabeled 1.0 mg MG Unlabeled, molecular grade 1.0 mg Phosphatase-labeled 0.1 mg Peroxidase-labeled 0.1 mg Fluorescein-labeled 0.5 mg E. coli O157:H7 Positive Control Antigen 1.0 ml Anti-Listeria, genus specific Unlabeled 1.0 mg Phosphatase-labeled 0.1 mg Peroxidase-labeled 0.1 mg Fluorescein-labeled 0.5 mg Listeria, genus specific Positive Control Antigen 1.0 ml Anti-Campylobacter, genus specific Unlabeled 1.0 mg Phosphatase-labeled 0.1 mg Peroxidase-labeled 0.1 mg Fluorescein-labeled 0.5 mg Campylobacter jejuni Positive Control Antigen 1.0 ml Anti-Borrelia burgdorferi Unlabeled 1.0 mg Phosphatase-labeled 0.1 mg Peroxidase-labeled 0.1 mg Fluorescein-labeled 0.5 mg Anti-Borrelia species Unlabeled 1.0 mg Peroxidase-labeled 0.1 mg Fluorescein-labeled 0.5 mg Borrelia burgdorferi Positive Control Antigen 1.0 ml Anti-Helicobacter pylori Unlabeled 1.0 mg Peroxidase-labeled 0.1 mg Fluorescein-labeled 0.5 mg Helicobacter pylori Positive Control Antigen 1.0 ml Anti-Renibacterium salmoninarum Unlabeled 1.0 mg Phosphatase-labeled 0.1 mg Peroxidase-labeled 0.1 mg Fluorescein-labeled 0.5 mg R. salmoninarun Positive Control Antigen 1.0 ml All BacTrace antibodies are made in goat. To order or for more information on KPL s line of protein detection products, contact us at / , FAX or visit us at ML For research use only. BacTrace is a registered trademark of KPL 2008 KPL, Inc. All rights reserved. BacTrace Anti-Salmonella (CSA) Broadly reactive; detects 23 Salmonella species Optimized for greatest specificity to Salmonella groups B,C,D and E Molecular grade antibody available for testing by PCR or other molecular methods. Salmonella typhimurium positive control available Anti-E.coli O157:H7 Highly specific; minimal crossreactivity to non-o157 serotypes of E.coli Molecular grade antibody available for testing by PCR or other molecular methods E.coli O157:H7 positive control available Gaithersburg, MD Phone: Fax: ISO 9001:2000 Registered

21 Peroxidase Conjugates KPL Peroxidase Conjugates: Time-tested, Sensitive and Reliable Since 1979 KPL has provided quality affinity purified antibodies to researchers worldwide. Over the years we have refined our production process to provide antibodies with high potency and consistent performance in immunoassays. From the start KPL gives careful consideration to immunogen preparation, using a highly purifed formulation to generate antiserum. KPL pools antiserum from multiple animals to reduce natural animal to animal serum variability. During the purification process our ISO 9001:2008-certified quality procedures impose rigorous standards for potency and cross-reactivity. 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 streptavidin. They are affinity purified and in some cases further adsorbed to minimize crossreactivity between animal species or shared reactivity with other immunoglobulin classes. HRP-labeled F(ab ) 2 fragment antibodies are offered for assays requiring extremely low background and absence of F(c)-mediated binding. KPL reacts HRP of the highest quality with affinity purified antibodies and streptavidin using the periodate method of Nakane and Kawaoi. Special features of HRP include: faster catalytic rate than alkaline phosphatase generates more product in shorter incubation times provides maximum sensitivity, low nonspecific binding ideal for ELISA, Western blotting and immunohistology applications.

22 Peroxidase Conjugates Peroxidase Conjugates Ordering Information (Partial listing) Catalog# Description Size HRP-labeled Goat Anti-Human IgG (H+L) 0.1 mg HRP-labeled Goat Anti-Human IgA+IgG+IgM (H+L), MSA 0.1 mg HRP-labeled Goat Anti-Human IgG (Fc) 0.1 mg HRP-labeled Goat Anti-Human IgG (γ) 1.0 mg HRP-labeled Goat Anti-Human IgM (μ) 1.0 mg HRP-labeled Goat Anti-Human IgE (ε) 1.0 mg HRP-labeled Goat Anti-Human IgG (H+L) 1.0 mg HRP-labeled Goat Anti-Human IgA+IgG+IgM (H+L) 1.0 mg HRP-labeled Goat Anti-Human IgA (α) 0.5 mg HRP-labeled F(ab ) 2 Goat Anti-Human IgG (γ) 0.5 mg HRP-labeled F(ab ) 2 Goat Anti-Human IgM (μ) 0.5 mg HRP-labeled F(ab ) 2 Goat Anti-Human IgG (H+L) 0.5 mg HRP-labeled Goat Anti-Human IgG (γ), Liquid 1.0 ml HRP-labeled Goat Anti-Human IgM (μ), Liquid 1.0 ml HRP-labeled Goat Anti-Human IgG (H+L), Liquid 1.0 ml HRP-labeled Goat Anti-Mouse IgG (H+L), HSA 0.1 mg HRP-labeled Goat Anti-Mouse IgG (H+L), RtSA, HSA 0.1 mg HRP-labeled Goat Anti-Mouse IgG (H+L), RbSA, HSA 0.1 mg HRP-labeled Goat Anti-Mouse IgG (γ), HSA 1.0 mg HRP-labeled Goat Anti-Mouse IgM (μ), HSA 1.0 mg HRP-labeled Goat Anti-Mouse IgG (H+L), HSA 1.0 mg HRP-labeled Goat Anti-Mouse IgG (H+L), XSA 1.0 mg HRP-labeled Goat Anti-Mouse IgA+IgG+IgM (H+L), HSA 1.0 mg HRP-labeled Goat Anti-Mouse IgG+IgM (H+L), HSA 1.0 mg HRP-labeled Goat Anti-Mouse IgA (α), HSA 0.5 mg HRP-labeled F(ab ) 2 Goat Anti-Mouse IgG (γ), HSA 0.5 mg HRP-labeled F(ab ) 2 Goat Anti-Mouse IgG (H+L), HSA 0.5 mg HRP-labeled Goat Anti-Mouse IgG (γ), HSA, Liquid 1.0 ml HRP-labeled Goat Anti-Mouse IgG (H+L), HSA, Liquid 1.0 ml HRP-labeled Goat Anti-Rabbit IgG (H+L) 1.0 mg HRP-labeled Goat Anti-Rabbit IgG (H+L), XSA 1.0 mg HRP-labeled Goat Anti-Rabbit IgG (H+L), HSA 1.0 mg HRP-labeled F(ab ) 2 Goat Anti-Rabbit IgG (H+L), HSA 0.5 mg HRP-labeled Goat Anti-Rabbit IgG (H+L), Liquid 1.0 ml HRP-labeled Goat Anti-Rabbit IgG (H+L), HSA, Liquid 1.0 ml HRP-labeled Streptavidin 0.5 mg HRP-labeled Streptavidin, Liquid, Molecular Grade 1.0 ml Visit our website at for a complete listing of HRP-labeled antibodies. To order or for more information on KPL s protein research products, contact us at / , FAX or visit us at 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. ELISA ABTS 1- and 2-Component Microwell Peroxidase Substrates SureBlue TM TMB Peroxidase Substrate SureBlue Reserve TM TMB Peroxidase Substrate TMB Peroxidase Substrate Blotting 4 CN Peroxidase Substrate TMB Membrane Peroxidase Substrate LumiGLO Chemiluminescent Substrate LumiGLO Reserve TM Chemiluminescent Substrate Whichever substrate you choose, enjoy the benefits of excellent signalto-noise and reproducibility. Gaithersburg, MD Phone: Fax: ISO 9001:2008 Registered For research use only KPL, Inc. All rights reserved. ABTS is a registered trademark of Roche Biochemicals. ML371-01

23 Ordering Information Catalog # Description Size Protein Detector ELISA Kits Each kit includes an AP- or HRP-labeled Conjugate, Coating Solution, Wash Solution Concentrate, 10% BSA Diluent/Blocking Solution Concentrate, Stop Solution and Substrate. Phosphatase AP ELISA Kit, Anti-Human 20 plates AP ELISA Kit, Anti-Rabbit and Anti-Mouse 20 plates Peroxidase HRP ELISA Kit, Anti-Human 20 plates HRP ELISA Kit, Anti-Mouse 20 plates HRP ELISA Kit, Anti-Rabbit 20 plates Substrates for ELISA Phosphatase Chromogenic Substrates BluePhos Microwell Substrate Kit 50 ml Same as above 600 ml Same as above 2.7 L FirePhos Microwell Substrate Kit 50 ml Same as above 200 ml Same as above 600 ml Same as above 2.7 L pnpp Phosphatase Substrate System 500 ml pnpp Phosphatase Tablets 100 tablets Peroxidase Chromogenic Substrates SureBlue TMB 1-Component Substrate 100 ml Same as above 400 ml Same as above 1 L SureBlue Reserve TMB 1-Component Substrate 100 ml Same as above 400 ml Same as above 1 L TMB 2-Component Substrate Kit 600 ml Same as above 2.7 L ABTS Microwell Substrate 1-Component 100 ml Same as above 600 ml Same as above 1 L ABTS Substrate System 2-Component 600 ml Same as above 2700 ml Peroxidase Chemiluminescent Substrates LumiGLO Chemiluminescent Substrate Kit 240 ml Same as above 720 ml Same as above 60 ml LumiGLO Reserve Chemiluminescent Substrate Kit 2400 cm Same as above 600 cm 2 Catalog # Description Size Phosphatase Chemiluminescent Substrates PhosphaGLO AP Substrate 30 ml Same as above 100 ml PhosphaGLO Reserve AP Substrate 30 ml Same as above 100 ml Antibody Conjugates All antibodies listed below are produced in goat. For a complete antibody listing, visit or refer to KPL's Product Catalog. Phosphatase ReserveAP Anti-Human IgG (H+L) 1.0 mg ReserveAP Anti-Mouse IgG (H+L), HSA 1.0 mg ReserveAP Anti-Rabbit IgG (H+L) 1.0 mg Peroxidase Anti-Human IgG (H+L) 1.0 mg Anti-Mouse IgG (H+L), HSA 1.0 mg Anti-Rabbit IgG (H+L) 1.0 mg Biotin-Labeled Anti-Human IgG (H+L) 0.5 mg Anti-Mouse IgG (H+L), HSA 0.5 mg Anti-Rabbit IgG (H+L) 0.5 mg Labeled Streptavidin AP-labeled 0.5 mg HRP-labeled 0.5 mg Assay Support Reagents Coating Solution Concentrate 50 ml Wash Solution Concentrate 800 ml Blocking Solutions % BSA Diluent/Blocking Solution 200 ml % BSA Diluent/Blocking Solution 1 L Milk Diluent/Blocking Solution 200 ml Conjugate Stabilizers APStabilizer 200 ml HRPStabilizer 200 ml Stop Solutions ABTS Stop Solution 200 ml APstop Solution 200 ml TMB Stop Solution 400 ml TMB Stop Solution 1 L TMB BlueSTOP Solution 400 ml TMB BlueSTOP Solution 1 L HSA: Human serum adsorbed ML To order, call or , fax , online at or contact your local KPL Sales Partner. ISO 9001:2000 Registered