This immunohistochemistry protocol describes the steps for performing the immunohistochemistry method with paraffin-embedded tissue sections.
Immunohistochemistry (IHC) is a method for demonstrating the presence and location of proteins within cells and in tissue context. In this experiment, a primary antibody binds specifically to a protein of interest present in a tissue.
Either chromogenic or fluorescent detection systems then visualize the antibody-antigen interaction. In chromogenic detection, an enzyme conjugated to the antibody cleaves a substrate to produce a coloured precipitate at the protein’s location, whereas, in fluorescent detection, a fluorophore is conjugated to the antibody and can be visualized using fluorescence microscopy.
IHC-P refers to the staining of tissues that have been fixed in neutral buffered formalin and then embedded in paraffin before being sectioned. This immunoassay is especially useful in diagnostics to determine morphological abnormalities and the presence of biomarkers indicative of the progression of diseases such as cancer.
This Inmunohistochemistry protocol contains:
- Materials and reactives
- Fixation and embedding of the tissue
- Cutting and mounting the section
- Deparaffinization and rehydration
- Antigen retrieval
- Immunohistochemical staining
- Counterstaining
- Dehydration and stabilization with mounting medium
- Troubleshooting
Related content
The complete guide to Inmunohistochemistry
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