Rat Angiotensin II ELISA Kit (A87284) employs the competitive enzyme immunoassay technique for the quantitative measurement of rat Angiotensin II in serum, plasma, tissue homogenates, and other biological fluids. The 96-well microtiter plate has been pre-coated with Angiotensin II antigen. During the incubation, Angiotensin II present in the samples or standards competes with the fixed amount of immobilized Angiotensin II for binding sites on the Biotinylated Anti-Angiotensin II Antibody. The more Angiotensin II present in a sample or standard, the less Biotinylated Anti-Angiotensin II Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Angiotensin II 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 Angiotensin II present in each sample or standard. The concentration of Angiotensin II can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.