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    tmb-elisa-substrate-high-sensitivity-ab171523.pdf

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TMB ELISA Substrate (High Sensitivity) (ab171523)

  • Datasheet
  • SDS
Submit a review Q&A (1)References (10)

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ELISA - TMB ELISA Substrate (High Sensitivity) (ab171523)
  • ELISA - TMB ELISA Substrate (High Sensitivity) (ab171523)

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Overview

  • Product name

    TMB ELISA Substrate (High 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 (High 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. ab171523 is not recommended for membrane or immunohistochemical applications that require a precipitating reaction product.

    Product Use:
    ab171523 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

    Shipped at Room Temperature. Store at +4°C. Store In the Dark.
  • Storage buffer

    pH: 3.50
    Constituents: 0.05% 3,3',5,5'-Tetramethylbenzidine, 79% Water, 0.1% Hydrogen peroxide
  • Concentration information loading...

Associated products

  • Related Products

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab171523 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 (High Sensitivity) (ab171523)
    ELISA - TMB ELISA Substrate (High Sensitivity) (ab171523)
    Rapid Detection
    TMB ELISA Substrate (High Sensitive) (ab171523) has fast enzyme kinetics for rapid color formation and improved assay sensitivity and performance.
  • ELISA - TMB ELISA Substrate (High Sensitivity) (ab171523)
    ELISA - TMB ELISA Substrate (High Sensitivity) (ab171523)

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 (10)

Publishing research using ab171523? Please let us know so that we can cite the reference in this datasheet.

ab171523 has been referenced in 10 publications.

  • Mantej J  et al. Autoantibodies to heat shock protein 60, 70, and 90 are not altered in the anti-SARS-CoV-2 IgG-seropositive humans without or with mild symptoms. Cell Stress Chaperones 26:735-740 (2021). PubMed: 34080135
  • Gao C  et al. SARS-CoV-2 Spike Protein Interacts with Multiple Innate Immune Receptors. bioRxiv N/A:N/A (2020). PubMed: 32766577
  • Silveria MA  et al. The Structure of an AAV5-AAVR Complex at 2.5 Å Resolution: Implications for Cellular Entry and Immune Neutralization of AAV Gene Therapy Vectors. Viruses 12:N/A (2020). PubMed: 33218165
  • McClean MCW  et al. A lineage-specific rapid diagnostic test (Chagas Sero K-SeT) identifies Brazilian Trypanosoma cruzi II/V/VI reservoir hosts among diverse mammalian orders. PLoS One 15:e0227828 (2020). PubMed: 31951634
  • Martinez Viedma MDP  et al. Peptide arrays incubated with three collections of human sera from patients infected with mosquito-borne viruses. F1000Res 8:1875 (2019). PubMed: 32201571
  • Liang W  et al. Structural mapping of hot spots within human CASPR2 discoidin domain for autoantibody recognition. J Autoimmun 96:168-177 (2019). PubMed: 30337146
  • Sánchez-Salguero E  et al. Infectious episodes during pregnancy, at particular mucosal sites, increase specific IgA1 or IgA2 subtype levels in human colostrum. Matern Health Neonatol Perinatol 5:9 (2019). PubMed: 31205733
  • Guo L  et al. A high-risk luminal A dominant breast cancer subtype with increased mobility. Breast Cancer Res Treat N/A:N/A (2019). PubMed: 30778902
  • Meneses GC  et al. Visceral leishmaniasis-associated nephropathy in hospitalised Brazilian patients: new insights based on kidney injury biomarkers. Trop Med Int Health 23:1046-1057 (2018). PubMed: 29987885
  • Morales Ruiz S  et al. Development of a lateral flow test for the rapid detection of Avibacterium paragallinarum in chickens suspected of having infectious coryza. BMC Vet Res 14:411 (2018). PubMed: 30567563

Customer reviews and Q&As

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Question

Inquiry: What is the ideal and maximum OD values after the TMB solution is stopped with phosphoric acid?

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Abcam community

Verified customer

Asked on Jun 02 2014

Answer

We recommend reading a stopped TMB solution around an OD of 2. The maximum OD will depend on your spectrophotometer, antibody concentration, and time.

Read More

Anja Hoffmann

Abcam Scientific Support

Answered on Jun 02 2014

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