Principle of Assay
Human Anti-Human VEGF Antibody ELISA Kit (A326372) is a 1h 30m indirect Enzyme-Linked Immunosorbent Assay (iELISA) designed for the in vitro quantitative determination of human Anti-Human VEGF Antibody in serum, plasma, cell culture supernatant, cell or tissue lysate, and other biological fluids. A 96-well microtiter plate has been pre-coated with assay-specific antigen. Standards and samples are added to the wells, where Anti-Human VEGF Antibody present in the samples binds to the immobilized antigen. Following incubation, unbound components are removed by washing and an HRP-conjugated detection antibody is added to the wells. This detection antibody binds the captured Anti-Human VEGF Antibody to form an indirect immune complex. After further incubation and washing, TMB substrate solution is added to visualize the HRP enzymatic reaction. This produces a blue-coloured product that changes to yellow following addition of acidic stop solution. The intensity of the yellow colour is directly proportional to the amount of Anti-Human VEGF Antibody captured in each well. The concentration of Anti-Human VEGF Antibody is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
