This immunoassay kit allows for the in vitro quantitative determination of Human 5-hydroxyindoleacetic acid,5-Hydroxyindoleacetic acid concentrations in serum, plasma, tissue homogenates, cell culture supernates, and other biological fluids.
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 5-Hydroxyindoleacetic acid, During the reaction, 5-Hydroxyindoleacetic acid in the sample or standard competes with a fixed amount of biotin-labeled 5-Hydroxyindoleacetic acid for sites on a pre-coated Monoclonal antibody specific to 5-Hydroxyindoleacetic acid. 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 5-Hydroxyindoleacetic acid in the samples is then determined by comparing the O.D. of the samples to the standard curve.