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Why should I block endogenous peroxidase acitivity
Some cells or tissues contain endogenous peroxidase. Using HRP conjugated antibody may result in high, non-specific background staining. This non-specific background can be significantly reduced by pre-treatment of cells/tissues with hydrogen peroxide prior to incubation with HRP conjugated antibody.
How do I know if my tissues contain endogenous peroxidase activity
Incubate your tissue sections with DAB substrate solution after rehydration to water. If it turned brown, the tissue contains endogenous peroxidase and a blocking step is needed.
What chemicals should I use to block endogenous peroxidase activity
Hydrogen peroxide (H2O2), a blocking agent in immunohistochemistry, is commonly used to block endogenous peroxidase acitivity. Pre-treatment with saturating amounts of hydrogen peroxide results in the irreversible inactivation of endogenous peroxidase.