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    tmb-elisa-substrate-fast-kinetic-rate-ab171524.pdf

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TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)

  • Datasheet
  • SDS
Submit a review Q&A (1)References (2)

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ELISA - TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
  • ELISA - TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
  • ELISA - TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)

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Overview

  • Product name

    TMB ELISA Substrate (Fast 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 (Fast 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. ab171524 is not recommended for membrane or immunohistochemical applications that require a precipitating reaction product.

    Product Use:
    ab171524 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:

    Product should be stored at 2-8°C. 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 short term (1-2 weeks). 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 (Slow Kinetic Rate) (ab171525)
    • 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 ab171524 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 (Fast Kinetic Rate) (ab171524)
    ELISA - TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
    Comparison of the kinetic curves of several TMB substrates at 500 pg/mL msIgG monitored over time at 650 nm.
  • ELISA - TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
    ELISA - TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
    Expanded graph at the lower concentrations using TMB ELISA Substrate (Fast Kinetic Rate) (ab171524) and TMB ELISA Substrate (Slowest Kinetic Rate) (ab171527). Both ab171524 and ab171527 have the same detection limit.
  • ELISA - TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
    ELISA - TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
    Comparison of the kinetic curves of TMB ELISA Substrate (Fast Kinetic Rate) (ab171524) at different concentrations of 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 ab171524? Please let us know so that we can cite the reference in this datasheet.

ab171524 has been referenced in 2 publications.

  • Mao SQ  et al. DNA G-quadruplex structures mold the DNA methylome. Nat Struct Mol Biol 25:951-957 (2018). PubMed: 30275516
  • Shen Z  et al. Delta-Like Ligand 4 Modulates Liver Damage by Down-Regulating Chemokine Expression. Am J Pathol 186:1874-1889 (2016). PubMed: 27171900

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Question


1-) What are the main differences between ab171524 (TMB ELISA Substrate - Fast Kinetic Rate) and ab171527 (TMB ELISA Substrate - Slowest Kinetic Rate)

2-) When should I choose using ab171524 or ab171527? I mean, is there any special situation (or condition) in which I should prefer one of these kit instead of the other?

Read More

Abcam community

Verified customer

Asked on Aug 21 2013

Answer



ab171524 (TMB ELISA Substrate - Fast Kinetic Rate) and ab171527 (TMB ELISA Substrate - Slowest Kinetic Rate) differ in the rate of color development and sensitivity.

TMB ELISA Substrate - Slowest Kinetic Rate has a relatively slower rate of color development and is a less sensitive substrate than ab171524 (TMB ELISA Substrate - Fast Kinetic Rate.

In general, one would select ab171527 (TMB ELISA Substrate - Slowest Kinetic Rate) if you feel the target protein detected in your assay is present in high amounts. The slow color development will allow you to stop the assay in a reasonable amount of time (eg. 5-10 minutes).

If you used ab171524 (TMB ELISA Substrate - Fast Kinetic Rate) as the substrate in the above case the color development may occur to quickly. This would lead to problems such as ODs quickly growing beyond the linear range of your spectrophotometer. In addition, while you are adding STOP solution to the first row of the plate, the remaining wells will continue to develop color while you add STOP solution to all the wells of the plate, leading to inconsistant results across the plate.

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Aug 21 2013

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