Principle of Assay
Human Thyroxine/T4 Antibody ELISA Kit (A78835) is a 3 hour indirect Enzyme-Linked Immunosorbent Assay (iELISA) designed for the in vitro quantitative determination of human Thyroxine/T4 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 Thyroxine/T4 Antibody binds to the immobilized antigen. Following incubation, unbound components are removed by washing and an enzyme-conjugated detection antibody is added to the wells. This detection antibody binds the captured Thyroxine/T4 Antibody to form an indirect immune 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 Thyroxine/T4 Antibody present in the sample. The concentration of Thyroxine/T4 Antibody is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
