Principle of Assay
Porcine Von Willebrand Factor ELISA Kit (A2062) employs the competitive enzyme immunoassay technique for the quantitative measurement of porcine Von Willebrand Factor in plasma or other biological fluids. The 96-well microtiter plate has been pre-coated with Von Willebrand Factor antigen. During the incubation, Von Willebrand Factor present in the samples or standards competes with the fixed amount of immobilized Von Willebrand Factor for binding sites on the Biotinylated Anti-Von Willebrand Factor Antibody. The more Von Willebrand Factor present in a sample or standard, the less Biotinylated Anti-Von Willebrand Factor Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Von Willebrand Factor 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 Von Willebrand Factor present in each sample or standard. The concentration of Von Willebrand Factor can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
