Principle of Assay
Canine Cholecystokinin ELISA Kit (A7107) employs the competitive enzyme immunoassay technique for the quantitative measurement of canine Cholecystokinin in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. The 96-well microtiter plate has been pre-coated with Cholecystokinin antigen. During the incubation, Cholecystokinin present in the samples or standards competes with the fixed amount of immobilized Cholecystokinin for binding sites on the Biotinylated Anti-Cholecystokinin Antibody. The more Cholecystokinin present in a sample or standard, the less Biotinylated Anti-Cholecystokinin Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Cholecystokinin Antibody is removed by washing, and an HRP-Avidin conjugate is added to the wells and the microtiter plate is incubated. 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 Cholecystokinin present in each sample or standard. The concentration of Cholecystokinin can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
