DRG 25-OH Vitamin D (Total) Elisa Kit

The DRG 25-OH Vitamin D (total) ELISA is a solid phase enzyme-linked immunosorbent assay (ELISA) based on the principle of competitive binding. The microtiter wells are coated with a monoclonal (mouse) antibody directed towards a unique antigenic site of the 25-OH Vitamin D (25-OH D) molecule. The patient sample is incubated together with Release Reagent in the wells to dissociate endogenous 25-OH D from Vitamin D binding protein (VDBP). Released 25-OH D then binds to the coated antibody of the well. After a washing step, biotin-labeled 25-OH D (Enzyme Conjugate) and peroxidase-labeled streptavidin (Enzyme Complex) are added. Added Biotin-25-OH D competes with endogenous 25-OH D for the binding to the coated antibody. Bound Biotin − 25-OH Vitamin D is then detected by Streptavidin-HRP. After incubation, unbound components are washed off. The amount of bound biotin-streptavidin complex is inversely proportional to the concentration of 25-OH Vitamin D in the sample. Subsequently, substrate solution is added and the color development is stopped after a defined time. The intensity of the color formed is inversely proportional to the 25-OH D concentration in the sample. The absorbance is measured at 450 nm with a microtiter plate reader.