IHC Troubleshooting: Non: specific Staining

The good immunohistochemical staining is a foundation of correct judgment for staining, since there are many steps and links through the whole immunohistochemistry process, each step and links are likely to affect the final result of the staining, so to do an immunohistochemistry quality slice is not a very easy thing.

There are often the incorrect signals except the correct positive signals in immunohistochemical staining, these incorrect staining called non-specific staining, There are a variety of reasons resulting non-specific staining. Summarized as follows:

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