This kit is based on Competitive-ELISA detection method and takes 2h assay time. The microplate provided in this kit has been precoated with Free HCY. Free HCY in the sample or standard competes with a fixed amount of biotin-labeled Free HCY for sites on a pre-coated antibody specific to Free HCY. Free components are washed off. After adding HRP-Streptavidin Conjugate (SABC), biotin specifically binds with streptavidin to form immune complexes. Wash off unbound conjugate. Add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Compare OD450 value of sample with the standard curve through curve fitting software to determine the concentration of Free HCY in the sample. The concentration of the target substance is inversely proportional to the OD450 value.
