The detection system builds on the secondary. It is now commonly used as multiple secondaries can bind to a single primary to amplify the staining intensity. Next, secondary antibodies bind to the primary antibody. Monoclonal antibodies have an affinity to only one epitope and tend to produce cleaner, more specific staining but are less sensitive or intense. Polyclonal antibodies have an affinity with, and bind to, multiple epitopes (or parts) or the target antigen, and as such are more prone to cross-react to non-target antigens. There are two main types of antibody, polyclonal and monoclonal. The first stage of IHC is the application of a primary antibody that binds specifically to the target antigen. Often, a pathologist will use a “panel” of multiple antigens to help fully classify a particular tumor. There are many hundreds of antigens that have been found to be diagnostically useful. Pathologists look for the presence or absence of particular antigens to assist with diagnosis. Target AntigenĪntigens are proteins that are within or on the surface of a cell. IHC is used as a diagnostic tool to assist in the diagnosis of solid tumors and cytological specimens and has been used as a mainstream diagnostic tool for almost half a century. Where H&E and Special Stains are non-specific, IHC is directed to a specific protein marker or markers. IHC has evolved to complement the Hematoxylin & Eosin (H&E) and Special Stain techniques that typically show tissue morphology (structure). IHC is used in histology to detect the presence of specific protein markers that can assist with accurate tumor classification and diagnosis.
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