Principle of Assay
This Human 5-MTHF ELISA Kit is an in vitro competitive enzyme-linked immunosorbent assay for the quantitative measurement of 5-MTHF.
A microplate has been precoated with 5-MTHF. Samples, standards and Anti-5-MTHF Antibody (Biotin) are added to the wells, where 5-MTHF in the sample competes with the fixed 5-MTHF to bind to the antibody. The wells are washed to remove unbound material and HRP-conjugated streptavidin is added. After incubation, the wells are washed again and TMB substrate is added. The TMB substrate is catalyzed by HRP to produce blue colouration. The reaction is terminated by addition of Stop Solution which stops the colour development and produces a colour change from blue to yellow. The intensity of the signal is inversely proportional to the amount of 5-MTHF in the sample and the intensity is measured at 450 nm.
