Principle of Assay
Prostaglandin D2 ELISA Kit (A326980) employs the competitive enzyme immunoassay technique for the quantitative measurement of Prostaglandin D2 in serum, plasma, and other biological fluids. The 96-well microtiter plate has been pre-coated with Prostaglandin D2 antigen. During the incubation, Prostaglandin D2 present in the samples or standards competes with the fixed amount of immobilized Prostaglandin D2 for binding sites on the Biotinylated Anti-Prostaglandin D2 Antibody. The more Prostaglandin D2 present in a sample or standard, the less Biotinylated Anti-Prostaglandin D2 Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Prostaglandin D2 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 Prostaglandin D2 present in each sample or standard. The concentration of Prostaglandin D2 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
