Immunohistochemistry (IHC) is used to Detect the Presence of a Specific Protein Marker that can Help with Tumour Categorization and Diagnosis


In histology, Immunohistochemistry (IHC) is used to detect the presence of certain protein markers that can help with tumor categorization and diagnosis. Hematoxylin and Eosin (H&E) and Special Stain methods, which normally display tissue morphology, have been supplemented by immunohistochemistry (structure). IHC is directed to a specific protein marker or markers, unlike H&E and Special Stains, which are non-specific. IHC has been used as a standard diagnostic method for about half a century to aid in the diagnosis of solid tumors and cytological materials.

Target Antigen

Antigens are proteins found inside or on the outside of a cell. To aid in the diagnosis, pathologists search for the presence or lack of specific antigens. Antigens have been found to be a diagnostically valuable number in the hundreds. A pathologist will frequently utilise a "panel" of numerous antigens to fully classify a tumor.

Primary Antibody

The application of a primary antibody that binds selectively to the target antigen is the first step in Immunohistochemistry (IHC). Polyclonal and monoclonal antibodies are the two main types of antibodies. Polyclonal antibodies have an affinity for many epitopes (or portions) of the target antigen and bind to them, making them more likely to cross-react with non-target antigens. Monoclonal antibodies bind to a single epitope and provide cleaner, more specific staining, although they are less sensitive and intense.

Secondary Antibody

The primary antibody is then bound by secondary antibodies. This is referred to as indirect  Immunohistochemistry (IHC). Multiple secondaries can attach to a single primary to increase the staining intensity, hence it's now widely employed.

The Detection System

The detecting system is built on top of the secondary system. Horseradish Peroxidase (HRP), for example, is conjugated (attached) to an antibody in modern chromogenic detection. Polymers are several enzymes coupled to an antibody that create more strong staining since there are more molecules for the chromogen to adhere to.


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