The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General NT, During the reaction, General NT in the sample or standard competes with a fixed amount of biotin-labeled General NT for sites on a pre-coated Monoclonal antibody specific to General NT. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of General NT in the samples is then determined by comparing the O.D. of the samples to the standard curve.