How does TMB react with HRP?
TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP, and the addition of an appropriate stop solution gives a clear yellow color that absorbs at 450nm.
How does TMB work?
Background: TMB (3,3′, 5,5”-tetramethylbenzidine) is used as a substrate to generate detectable signal in ELISA. The reaction between the TMB substrate and a peroxidase, typically horseradish peroxidase (HRP), produces a measur- able color change that correlates with analyte level.
Is TMB a substrate for HRP?
3,3’5,5′-Tetramethylbenzidine (TMB) is a safe-sensitive substrate used for the detection of Horseradish Peroxidase (HRP) activity. It yields a blue colour (Amax = 652nm) and changes to yellow (Amax = 450nm) upon the addition of either sulfuric or phosphoric acid.
Why HRP is used in ELISA?
HRP is used in immunohistochemistry and ELISA because it generates colored compounds. For detection of an antigen or protein molecule, HRP substrates have been designed so that they will generate a chemiluminescent, chromogenic, or fluorescent signal upon oxidation.
What is HRP and TMB ELISA?
TMB (3,3′,5,5′-Tetramethylbenzidine) is a substrate for horseradish peroxidase (HRP), a common label conjugated to antibodies for applications such as immunohistochemistry, western blotting, and ELISA. The combination of HRP and hydrogen peroxide causes the oxidation of TMB, resulting in the formation of a blue color.
What is HRP substrate?
Horseradish peroxidase (HRP) is an enzyme used to amplify signal in photometric assays by catalyzing the conversion of chromogenic or chemiluminescent substrates for the detection of targets such as proteins, carbohydrates, and nucleic acids.
Does TMB contain H2O2?
Product Description: TMB No Hydrogen Peroxide One Component Substrate contains 3,3′,5,5′-tetramethylbenzidine in a mildly acidic buffer.
What does HRP do in ELISA?