Principle of Assay
Chloramphenicol ELISA Test Kit (A327188) employs the competitive enzyme immunoassay technique for the quantitative measurement of Chloramphenicol in fish, shrimp, eggs, honey, meat (beef, chicken and pork), milk, milk powder, condensed milk, cell lysis solution and serum. The 96-well microtiter plate has been pre-coated with Chloramphenicol. During the incubation, Chloramphenicol present in the samples or standards competes with the fixed amount of immobilized Chloramphenicol for binding sites of the Anti-Chloramphenicol Antibody, a HRP-Conjugated Secondary Antibody is added to the wells at the same time. The more Chloramphenicol present in a sample or standard, the less Anti-Chloramphenicol 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 Chloramphenicol present in each sample or standard. The concentration of Chloramphenicol can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
