Description
Principle of the Assay: This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Glucagon has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Glucagon present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for Glucagon is added to the wells and binds to the combination of capture antibody-Glucagon in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of Glucagon present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven Glucagon standard dilutions and Glucagon sample concentration determined: