It is said that 0.5M H2SO4 acts as a stop solution to stop the reaction and turns the green color to yellow.I would like to know how it halts the reaction and how TMB and HRP play their role in the reaction in a bioassay.
Thank you,
Sven Buelow
H2SO4 makes the solution acidic and thus inactivates horseradish peroxidase, stopping the enzymatic reaction. It also stabilizes the colorful product of the reaction.
TMB is the hydrogen donor for the reduction of H2O2 to water, in the reaction, TMB is converted to its blue counterpart (see link).
https://en.wikipedia.org/wiki/3,3',5,5'-Tetramethylbenzidine
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