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    tmb-elisa-substrate-highest-sensitivity-ab171522.pdf

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TMB ELISA Substrate (Highest Sensitivity) (ab171522)

  • Datasheet
  • SDS
Submit a review Q&A (4)References (6)

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ELISA - TMB ELISA Substrate (Highest Sensitivity) (ab171522)
  • ELISA - TMB ELISA Substrate (Highest Sensitivity) (ab171522)

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Overview

  • Product name

    TMB ELISA Substrate (Highest Sensitivity)
    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 (Highest Sensitive) 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. ab171522 is not recommended for membrane or immunohistochemical applications that require a precipitating reaction product.

    Product Use:
    ab171522 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 (High Sensitivity) (ab171523)
    • TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
    • 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 ab171522 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
Use at an assay dependent concentration.
Notes
ELISA
Use at an assay dependent concentration.
Application notes
Is unsuitable for IHC-Fr or IHC-P.

Images

  • ELISA - TMB ELISA Substrate (Highest Sensitivity) (ab171522)
    ELISA - TMB ELISA Substrate (Highest Sensitivity) (ab171522)
    Rapid Detection
    TMB ELISA Substrate (Highest Sensitive) (ab171522) has the fastest overall enzyme kinetics for rapid color formation and improved assay sensitivity and performance.
  • ELISA - TMB ELISA Substrate (Highest Sensitivity) (ab171522)
    ELISA - TMB ELISA Substrate (Highest Sensitivity) (ab171522)

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 (6)

Publishing research using ab171522? Please let us know so that we can cite the reference in this datasheet.

ab171522 has been referenced in 6 publications.

  • Biferi MG  et al. Systemic Treatment of Fabry Disease Using a Novel AAV9 Vector Expressing a-Galactosidase A. Mol Ther Methods Clin Dev 20:1-17 (2021). PubMed: 33335943
  • Li X  et al. The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats. Clin Transl Med 9:16 (2020). PubMed: 32056048
  • Chernyavtseva A  et al. Differential recognition of peptides within feline coronavirus polyprotein 1 ab by sera from healthy cats and cats with feline infectious peritonitis. Virology 532:88-96 (2019). PubMed: 31048107
  • Bailey JK  et al. Synthesis of multivalent glycopeptide conjugates that mimic an HIV epitope. Tetrahedron 72:6091-6098 (2016). PubMed: 28190897
  • Mathesz A  et al. [Integrated optical sensor for rapid analysis of biological samples]. Orv Hetil 156:2116-9 (2015). PubMed: 26686748
  • Hersey JS  et al. Functionalized Nanofiber Meshes Enhance Immunosorbent Assays. Anal Chem 87:11863-70 (2015). PubMed: 26551162

Customer reviews and Q&As

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

1-4 of 4 Abreviews or Q&A

Question

Why do you offer multiple TMB substrates for microwell applications?

Read More

Abcam community

Verified customer

Asked on May 14 2013

Answer

Our TMB substrates have the same lower detection limit but the different formulations provide differing rates of color formation at all analyte levels. This provides flexibility in the timing of your assay.

Read More

Abcam Scientific Support

Answered on May 14 2013

Question

Will the TMB substrates freeze when stored at 2oC to 8oC

Read More

Abcam community

Verified customer

Asked on May 14 2013

Answer

No.

Read More

Abcam Scientific Support

Answered on May 14 2013

Question

What solvent is used in TMB Substrates?

Read More

Abcam community

Verified customer

Asked on May 14 2013

Answer

A proprietary non-toxic, non-hazardous, non-carcinogenic solvent is used in our TMB Substrates.

Read More

Abcam Scientific Support

Answered on May 14 2013

Question

How can the intensity of the TMB reaction be reduced or slowed?

Read More

Abcam community

Verified customer

Asked on May 14 2013

Answer

To reduce the intensity of a reaction, it is recommended that the antibodies or conjugates be diluted.

Read More

Abcam Scientific Support

Answered on May 14 2013

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