The fT3 test is a solid phase competitive enzyme immunoassay. Patientserum samples, standards, and T3-Enzyme Conjugate Working Reagentis added to wells coated with monoclonal T3 antibody. fT3 in the patientspecimen and the T3 labeled conjugate compete for available binding siteson the antibody. After a 60 minutes incubation at room temperature, thewells are washed with water to remove unbound T3 conjugate. A solutionof H2O2/TMB is then added and incubated for 20 minutes, resulting inthe development of blue color. The color development is stopped with theaddition of 3N HCl, and the absorbance is measured spectrophotometricallyat 450 nm. The intensity of the color formed is proportional to the amountof enzyme present and is inversely related to the amount of unlabeled fT3in the sample
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