Principle of Assay
Ochratoxin A ELISA Test Kit (A327236) employs the competitive enzyme immunoassay technique for the quantitative measurement of Ochratoxin A in cereals, feed, fish and shrimp. The 96-well microtiter plate has been pre-coated with Ochratoxin A. During the incubation, Ochratoxin A present in the samples or standards competes with the fixed amount of immobilized Ochratoxin A for binding sites of the Anti-Ochratoxin A Antibody, a HRP-Conjugated Secondary Antibody is added to the wells at the same time. The more Ochratoxin A present in a sample or standard, the less Anti-Ochratoxin A 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 Ochratoxin A present in each sample or standard. The concentration of Ochratoxin A can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
