This TAT enzyme linked immunosorbent assay applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific for TAT. Standards or samples are then added to the microtiter plate wells and TAT if present, will bind to the antibody pre-coated wells. In order to quantitatively determine the amount of TAT present in the sample, a standardized preparation of horseradish peroxidase (HRP) -conjugated polyclonal antibody, specific for TAT are added to each well to sandwich the TAT immobilized on the plate. The microtiter plate undergoes incubation, and then the wells are thoroughly washed to remove all unbound components. Next, A and B substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period.Only those wells that contain TAT and enzyme-conjugated antibody will exhibit a change in colour. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the colour change is measured spectrophotometrically at a wavelength of 450 nm.
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