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Non-specific background in immunohistochemistry

Explore causes for non-specific background in immunohistochemistry.

Why is there a non-specific background?

Possible causes

Solution

Antibody binding non-specifically to tissue or FC receptors.

  • Reduce non-specific binding by pre-treatment of the sample with hydrogen peroxide before incubation with HRP-conjugated antibody.

    Variations:

    For fragile samples, where presevation of the morphology is required, dilute the hydrogen peroxide in methanol.

    For cell surface or membrane proteins use hydrogen peroxide in PBS or quench with peroxide after first incubating sections or cells with the primary antibody.