Principle of Assay
Reverse Triiodothyronine ELISA Kit (A3809) employs the competitive enzyme immunoassay technique for the quantitative measurement of Reverse Triiodothyronine in serum, plasma or other biological fluids. The 96-well microtiter plate has been pre-coated with Reverse Triiodothyronine antigen. During the incubation, Reverse Triiodothyronine present in the samples or standards competes with the fixed amount of immobilized Reverse Triiodothyronine for binding sites on the Biotinylated Anti-Reverse Triiodothyronine Antibody. The more Reverse Triiodothyronine present in a sample or standard, the less Biotinylated Anti-Reverse Triiodothyronine Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Reverse Triiodothyronine Antibody is removed by washing, and an HRP-Avidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is inversely proportional to the amount of Reverse Triiodothyronine present in each sample or standard. The concentration of Reverse Triiodothyronine can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
