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Practical aspects of Ab assays Each Ab assay has its own characteristic advantages and disadvantages based on technical difficulty, intra- and interassay variability (i.e., precision), sensitivity for measuring Abs, specificity for the target protein or ligand, susceptibility to false-positive or false-negative results, and cost.
ELISA is a popular format of wet-lab type analytic biochemistry assay that uses a solid-phase enzyme immunoassay (EIA) to detect the presence of a substance, usually an antigen, in a liquid sample or wet sample. The secondary antibody, specific to the primary antibody, is added. 2  This second antibody is coupled to the enzyme. 3  A substrate is added, and remaining enzymes elicit a chromogenic or fluorescent signal. 4  The reaction is stopped to prevent eventual saturation of the signal.