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Immunochemical techniques include: enzyme-linked immunosorbent assay, immunoblotting (e.g., Western blot assay), precipitation and agglutination reactions, immunoelectrophoresis, immunophenotyping, immunochromatographic assay and cyflometry. One of the earliest examples of immunochemistry is the Wasserman test to detect syphilis.
Immunodiffusion is a laboratory technique used to detect and quantify antigens and antibodies by observing their interactions within a gel medium. [1] This technique involves the diffusion of antigens and antibodies through a gel, usually agar, resulting in the formation of a visible precipitate when they interact.
In the zone of equivalence, the formation of precipitin complexes is optimal. Extensive lattices of antigen and antibody are formed by cross-linking. At high concentrations of antigen, the average size of antibody-antigen complexes is once again small because few antibody molecules are available to cross-link antigen molecules together.
Therefore, precipitation near the center of the circle is usually less dense than it is near the circle's outer edge, where antigen is less concentrated. [ 1 ] [ 2 ] Expansion of the circle reaches an endpoint and stops when free antigen is depleted and when antigen and antibody reach equivalence.
Immunoprecipitation of intact protein complexes (i.e. antigen along with any proteins or ligands that are bound to it) is known as co-immunoprecipitation (Co-IP). Co-IP works by selecting an antibody that targets a known protein that is believed to be a member of a larger complex of proteins.
Precipitation occurs with most antigens because the antigen is multivalent (i.e. has several antigenic determinants per molecule to which antibodies can bind). Antibodies have at least two antigen binding sites (and in the case of immunoglobulin M there is a multimeric complex with up to 10 antigen binding sites), thus large aggregates or gel ...
In addition to the binding of an antibody to its antigen, the other key feature of all immunoassays is a means to produce a measurable signal in response to the binding. Most, though not all, immunoassays involve chemically linking antibodies or antigens with some kind of detectable label.
Immunocytochemistry labels individual proteins within cells, such as TH (green) in the axons of sympathetic autonomic neurons.. Immunocytochemistry (ICC) is a common laboratory technique that is used to anatomically visualize the localization of a specific protein or antigen in cells by use of a specific primary antibody that binds to it.