Principle of Assay
Crystal Violet ELISA Test Kit (A327196) employs the competitive enzyme immunoassay technique for the quantitative measurement of Crystal Violet in water, fish, shrimp. The 96-well microtiter plate has been pre-coated with Crystal Violet. During the incubation, Crystal Violet present in the samples or standards competes with the fixed amount of immobilized Crystal Violet for binding sites of the Anti-Crystal Violet Antibody, a HRP-Conjugated Secondary Antibody is added to the wells at the same time. The more Crystal Violet present in a sample or standard, the less Anti-Crystal Violet Antibody that binds to the plate. Following incubation, unbound HRP-Conjugated Secondary Antibody is removed by washing. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is inversely proportional to the amount of Crystal Violet present in each sample or standard. The concentration of Crystal Violet can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
