Principle of Assay
Mouse Proteinase-3 Anti-Neutrophil Cytoplasmic Antibody ELISA Kit (A303541) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of mouse Proteinase-3 Anti-Neutrophil Cytoplasmic 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 Proteinase-3 Anti-Neutrophil Cytoplasmic 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 Proteinase-3 Anti-Neutrophil Cytoplasmic 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 Proteinase-3 Anti-Neutrophil Cytoplasmic Antibody captured in each well. The concentration of Proteinase-3 Anti-Neutrophil Cytoplasmic Antibody is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
