Principle of the Procedure

This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre coated with Human 25 OH D. Standards or samples are added to the appropriate microtiter plate wells then with a biotin conjugated antibody specific to Human 25 OH D. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme substrate reaction is terminated by the a ddition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human 25 OH D in the samples is then determined by comparing the OD of the samples to the standard curve.