Principle of Assay
Rat Corticosterone ELISA Kit (A327154) employs the competitive enzyme immunoassay technique for the quantitative measurement of rat Corticosterone in serum, plasma, urine, and saliva. The 96-well microtiter plate has been pre-coated with Corticosterone antigen. During the incubation, Corticosterone present in the samples or standards competes with the fixed amount of immobilized Corticosterone for binding sites on the Biotinylated Anti-Corticosterone Antibody. The more Corticosterone present in a sample or standard, the less Biotinylated Anti-Corticosterone Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Corticosterone Antibody is removed by washing, and an HRP-Streptavidin 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 Corticosterone present in each sample or standard. The concentration of Corticosterone can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
