Description
This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for Glucagon has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any Glucagon present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-conjugated antibody specific for Glucagon is added to each well and incubate. Following a washing to remove unbound substances, streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Glucagon bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm ±2nm.The concentration of Glucagon in the sample is then determined by comparing the O.D of samples to the standard curve