Principle of Assay
Mouse Antithrombin III/ATIII Antibody ELISA Kit (A76173) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of mouse Antithrombin III/ATIII 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 Antithrombin III/ATIII 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 Antithrombin III/ATIII 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 Antithrombin III/ATIII Antibody captured in each well. The concentration of Antithrombin III/ATIII Antibody is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
