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    tmb-elisa-substrate-slow-kinetic-rate-ab171525.pdf

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TMB ELISA Substrate (Slow Kinetic Rate) (ab171525)

  • Datasheet
  • SDS
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ELISA - TMB ELISA Substrate (Slow Kinetic Rate) (ab171525)

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    Overview

    • 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

    • 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

    • 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
    ELISA
    1/1. 100 µL of substrate solution is added to each well.
    Notes
    ELISA
    1/1. 100 µL of substrate solution is added to each well.
    Application notes
    Is unsuitable for IHC-Fr or IHC-P.

    Images

    • ELISA - TMB ELISA Substrate (Slow Kinetic Rate) (ab171525)
      ELISA - TMB ELISA Substrate (Slow Kinetic Rate) (ab171525)
      Comparison of the kinetic curves of several TMB substrates at 500 pg/mL msIgG monitored over time at 650 nm.

    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.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (2)

    Publishing research using ab171525? Please let us know so that we can cite the reference in this datasheet.

    ab171525 has been referenced in 2 publications.

    • Zhang X  et al. Chemical profiling of DNA G-quadruplex-interacting proteins in live cells. Nat Chem 13:626-633 (2021). PubMed: 34183817
    • Gaudreault NN  et al. SARS-CoV-2 infection, disease and transmission in domestic cats. Emerg Microbes Infect 9:2322-2332 (2020). PubMed: 33028154

    Customer reviews and Q&As

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