Principle of Assay
Human Anti-Vinculin Antibody ELISA Kit (A303085) is a 3 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Anti-Vinculin Antibody in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for Anti-Vinculin Antibody. Standards and samples are added to the wells, where Anti-Vinculin Antibody binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and an HRP-conjugated detection antibody specific for Anti-Vinculin Antibody is added to the wells. The detection antibody binds the captured Anti-Vinculin Antibody to form an antibody-antigen-antibody sandwich 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-Vinculin Antibody captured in each well. The concentration of Anti-Vinculin Antibody is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
