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AAT Bioquest

What should I do to achieve the optimal results for my blocking experiments for immunohistochemistry (IHC)?

Posted May 2, 2024


Answer

These are some things you can do to achieve optimal results for your blocking experiments for immunohistochemistry (IHC):

  • Make sure to select a blocking buffer that provides the highest signal-to-noise ratio. 
  • Monitor signal strength (positive control) as well as background (negative control) using different blocking reagents. 
  • Maintain optimal assay conditions by using the same blocking buffer for both, antibody dilution and blocking steps. 
  • Ensure there are no substances in the blocking buffer that may interfere with a particular assay. For example, the biotin content makes non-fat dry milk inappropriate for use with any detection system that includes a biotin-binding protein.
Additional resources

Technical aspects of immunohistochemistry

Immunohistochemistry (IHC)

Stayright™ Purple HRP Staining Kit