Principle of Assay
Acrylamide-ES ELISA Test Kit (A327173) employs the competitive enzyme immunoassay technique for the quantitative measurement of Acrylamide-ES in food samples such as potato/corn chips, french fries, cereals, and breads. The 96-well microtiter plate has been pre-coated with Acrylamide-ES. During the incubation, Acrylamide-ES present in the samples or standards competes with the fixed amount of immobilized Acrylamide-ES for binding sites of the Anti-Acrylamide-ES Antibody, a HRP-Conjugated Secondary Antibody is added to the wells at the same time. The more Acrylamide-ES present in a sample or standard, the less Anti-Acrylamide-ES 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 Acrylamide-ES present in each sample or standard. The concentration of Acrylamide-ES can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
