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A sensitive technique for accurately determining specific molecules in a mixed sample. The amount of protein or other antigen in a given sample is determined by means of an enzyme-catalyzed color change, avoiding both the hazards and expense of radioactive techniques. It takes various forms. In the most common form, two antibody preparations are used in ELISA. An antibody (primary) specific to the test protein is adsorbed onto a solid substrate, and a known amount of the sample is added; all molecules of the test protein in the sample are bound by the antibody. A second antibody specific for a second site on the test protein is added; this is conjugated with an enzyme, which catalyses a colour change in the fourth reagent, added finally. The color change is measured photometrically and compared against a standard curve to give the concentration of protein in the sample. ELISA is widely used for diagnostic and other purposes.
Publication Source: Food and Agricultural Organization of the United Nations (FAO)
Publication Date: 1999
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