The DRG Hepcidin-25 ELISA Kit 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 an antigenic site of the Hepcidin-25 molecule. Endogenous Hepcidin-25 of a specimen sample competes with a Hepcidin-25-biotin conjugate for binding to the coated antibody.
After incubation, the unbound conjugate is washed off and a streptavidin-peroxidase enzyme complex is added to each well. After incubation, unbound enzyme complex is washed off and substrate solution is added. The blue colour development is stopped after a short incubation time, turning the colour from blue to yellow. The intensity of colour developed is reverse proportional to the concentration of Hepcidin in the specimen sample.