Immunohistochemistry Application Guide

How immunohistochemistry works

Antibody binding is visualised in a tissue section using an enzyme-based chromogenic system or fluorescent detection. Tissue processing and antigen retrieval can change epitope accessibility and therefore the observed staining pattern.

1. Select and prepare frozen or fixed tissue sections.
2. Optimise antigen retrieval for the fixation method and target.
3. Block endogenous activity and non-specific binding.
4. Stain, detect, counterstain and interpret against appropriate controls.

Experimental planning and controls

Plan positive and negative tissue controls where possible and record fixation, retrieval, antibody concentration and detection chemistry. Expected tissue distribution and subcellular localisation should inform interpretation, but staining should be considered alongside control performance.