Principle of Assay
Porcine Apelin ELISA Kit (A76014) employs the competitive enzyme immunoassay technique for the quantitative measurement of porcine Apelin in serum, plasma, tissue homogenates, and other biological fluids. The 96-well microtiter plate has been pre-coated with Apelin antigen. During the incubation, Apelin present in the samples or standards competes with the fixed amount of immobilized Apelin for binding sites on the Biotinylated Anti-Apelin Antibody. The more Apelin present in a sample or standard, the less Biotinylated Anti-Apelin Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Apelin 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 Apelin present in each sample or standard. The concentration of Apelin can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
