Principle of Assay
Mouse Rheumatoid Factor IgA ELISA Kit (A247074) is a 3 hour indirect Enzyme-Linked Immunosorbent Assay (iELISA) designed for the in vitro quantitative determination of mouse Rheumatoid Factor IgA 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 Rheumatoid Factor IgA binds to the immobilized antigen. Following incubation, unbound components are removed by washing and an enzyme-conjugated detection antibody is added. This detection antibody binds the captured Rheumatoid Factor IgA to form an antigen-antibody-detection antibody sandwich complex. After further incubation and washing, TMB substrate solution is added to visualize the enzyme-mediated 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 Rheumatoid Factor IgA captured in each well. The concentration of Rheumatoid Factor IgA is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
