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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.
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.
Antigens are bound to antibodies through weak and noncovalent interactions such as electrostatic interactions, hydrogen bonds, Van der Waals forces, and hydrophobic interactions. [4] The principles of specificity and cross-reactivity of the antigen-antibody interaction are useful in clinical laboratory for diagnostic purposes.
Immunoelectrophoresis is a general term describing many combinations of the principles of electrophoresis and reaction of antibodies, also known as immunodiffusion. [1] Agarose as 1% gel slabs of about 1 mm thickness buffered at high pH (around 8.6) is traditionally preferred for electrophoresis and the reaction with antibodies. The agarose was ...
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.
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As more antigen is added, the amount of protein precipitated increases until the antigen/antibody molecules are at an optimal ratio. This is known as the zone of equivalence or equivalence point. When the amount of antigen in solution exceeds the amount of antibody, the amount of precipitation will decrease. This is known as the antigen excess ...