Troubleshooting - Non-specific staining

Immunohistochemistry (IHC) troubleshooting for no staining, high background and non-specific staining.

Problems Possible reasons Solutions
High background Insufficient washing after antibody incubation Wash with PBS appropriately
Antibody concentration was too high Decrease the antibody concentration
Presence of endogenous peroxidase Block endogenous peroxidase activity properly
The secondary antibody is as the same species as the tissue's origin Change the secondary antibody

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