Principle of Assay
Universal Smooth Muscle Actin ELISA Kit (A335457) employs the competitive enzyme immunoassay technique for the quantitative measurement of universal Smooth Muscle Actin in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The 96-well microtiter plate has been pre-coated with Smooth Muscle Actin antigen. During the incubation, Smooth Muscle Actin present in the samples or standards competes with the fixed amount of immobilized Smooth Muscle Actin for binding sites on the Biotinylated Anti-Smooth Muscle Actin Antibody. The more Smooth Muscle Actin present in a sample or standard, the less Biotinylated Anti-Smooth Muscle Actin Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Smooth Muscle Actin 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 Smooth Muscle Actin present in each sample or standard. The concentration of Smooth Muscle Actin can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
