TMB ELISA Substrate (Slow Kinetic Rate) (ab171525)
Overview
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Product name
TMB ELISA Substrate (Slow Kinetic Rate)
See all ELISA substrate reagents -
Tested applications
Suitable for: ELISAmore details
Unsuitable for: IHC-Fr or IHC-P -
General notes
Abcam’s TMB ELISA Substrate (Slow Kinetic Rate) detects horseradish peroxidase (HRP) activity and contains 3,3’,5,5’-tetramethylbenzidine in a mildly acidic buffer. The substrate is supplied as a ready to use solution. Unreacted substrate should be colorless or very light yellow in appearance. When this substrate system is reacted with peroxidase, a soluble blue reaction product is obtained. The reaction can be stopped using appropriate stop solution, producing a soluble yellow or soluble blue reaction product, depending upon the stop reagent used, which is stable for at least 1 hour. ab171525 is not recommended for membrane or immunohistochemical applications that require a precipitating reaction product.
Product Use:
ab171525 is supplied as a ready to use solution. The product should be allowed to equilibrate to room temperature (25°C) prior to use. For microwell applications, 100 µL of substrate solution is added to each well. A soluble blue reaction product develops which can be read at 370 nm or in a range of 620 nm to 650 nm. For best results, sample absorbance values should be monitored and read before absorbance values exceed 2.0 OD units. In endpoint assays, the substrate reaction can be stopped using equal volumes of 1 N HCl, 0.6 N sulfuric acid, or one of the stop solutions (ab171529 and ab171531). Addition of acid turns the blue color to yellow and stops the enzymatic reaction. In the case of the 650 nm Stop Solutions for TMB Substrate (ab171531), the blue color does not change. Since stopping the reaction increases sample absorbance values approximately three fold, unless using the 650 nm Stop Solutions for TMB Substrate (ab171531), sample OD values should be monitored and substrate reaction stopped when values reach approximately 0.7 OD units. After stopping with the 450 nm Stop Solutions for TMB Substrate (ab171529), a soluble yellow product develops which is read in the 450 nm range. Stopping with the 650 nm Stop Solutions for TMB Substrate (ab171531) produces a soluble blue product which is read in the 650 nm range. Dilution of the substrate is not recommended. To reduce the intensity of a reaction, it is recommended that the antibodies or conjugates be diluted.Storage Instructions:
Exposure to direct sunlight and other UV sources should be avoided due to the light sensitive nature of the TMB molecule.
Properties
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Form
Liquid -
Storage instructions
Store at +4°C. Please see notes section. -
Storage buffer
pH: 1
Constituents: 0.05% TMB, 79% Water, 0.1% Hydrogen peroxide -
Concentration information loading...
Associated products
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Related Products
- TMB ELISA Substrate (Highest Sensitivity) (ab171522)
- TMB ELISA Substrate (High Sensitivity) (ab171523)
- TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
- TMB ELISA Substrate (Slower Kinetic Rate) (ab171526)
- TMB ELISA Substrate (Slowest Kinetic Rate) (ab171527)
- 450 nm Stop Solution for TMB Substrate (ab171529)
- 650 nm Stop Solution for TMB Substrate (ab171531)
- Phosphate Buffered Saline (PBS) Casein ELISA Reagent (ab171532)
- Tris Buffered Saline (TBS) Casein ELISA Reagent (ab171533)
- Immunoassay Blocking Buffer (ab171534)
- Immunoassay Blocking (BSA Free) (ab171535)
- AP Stability Conjugate Reagent (ab171536)
- HRP Stability Conjugate Reagent (ab171537)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab171525 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ELISA |
1/1. 100 µL of substrate solution is added to each well.
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Notes |
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ELISA
1/1. 100 µL of substrate solution is added to each well. |
Images
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab171525 has been referenced in 4 publications.
- Bold D et al. Development of an Indirect ELISA for the Detection of SARS-CoV-2 Antibodies in Cats. Front Vet Sci 9:864884 (2022). PubMed: 35754530
- Zhang X et al. Chemical profiling of DNA G-quadruplex-interacting proteins in live cells. Nat Chem 13:626-633 (2021). PubMed: 34183817
- Cool K et al. Infection and transmission of ancestral SARS-CoV-2 and its alpha variant in pregnant white-tailed deer. bioRxiv N/A:N/A (2021). PubMed: 34426811
- Gaudreault NN et al. SARS-CoV-2 infection, disease and transmission in domestic cats. Emerg Microbes Infect 9:2322-2332 (2020). PubMed: 33028154