Principle of Assay
Mouse Protein Carbonyl ELISA Kit (A326804) employs the competitive enzyme immunoassay technique for the quantitative measurement of mouse Protein Carbonyl in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The 96-well microtiter plate has been pre-coated with Protein Carbonyl antigen. During the incubation, Protein Carbonyl present in the samples or standards competes with the fixed amount of immobilized Protein Carbonyl for binding sites on the Biotinylated Anti-Protein Carbonyl Antibody. The more Protein Carbonyl present in a sample or standard, the less Biotinylated Anti-Protein Carbonyl Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Protein Carbonyl 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 Protein Carbonyl present in each sample or standard. The concentration of Protein Carbonyl can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
