Principle of Assay
Adenosine Diphosphate ELISA Kit (A2051) employs the competitive enzyme immunoassay technique for the quantitative measurement of Adenosine Diphosphate in tissue homogenates, cell lysates or other biological fluids. The 96-well microtiter plate has been pre-coated with Adenosine Diphosphate antigen. During the incubation, Adenosine Diphosphate present in the samples or standards competes with the fixed amount of immobilized Adenosine Diphosphate for binding sites on the Biotinylated Anti-Adenosine Diphosphate Antibody. The more Adenosine Diphosphate present in a sample or standard, the less Biotinylated Anti-Adenosine Diphosphate Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Adenosine Diphosphate Antibody is removed by washing, and an HRP-Avidin 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 Adenosine Diphosphate present in each sample or standard. The concentration of Adenosine Diphosphate can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
