Principle of Assay
Human P53 Antibody ELISA Kit (A86966) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human P53 Antibody in serum, plasma, tissue homogenates, 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 P53 Antibody binds to the immobilized antigen. Following incubation, unbound components are removed by washing and a biotinylated antigen detection reagent is added to the wells. This reagent binds the captured P53 Antibody to form an antigen-antibody-antigen sandwich complex. After further incubation and washing, HRP-Streptavidin conjugate is added to the wells. TMB substrate solution is then 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 P53 Antibody captured in each well. The concentration of P53 Antibody is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
