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This assay employs the quantitative sandwich enzyme immunoassay technique. Amonoclonal antibody specific for human EGF has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any human EGF present isbound by the immobilized antibody. After washing away any unbound substances, apolyclonal antibody conjugated to horseradish peroxidase for human EGF is addedtothe wells. Following a wash to remove any unbound reagent, TMB substrate solution(Chromogenic agent) is added to the wells and color develops in proportion totheamount of human EGF bound in the initial step. The color development is stopped,and the intensity of the color is measured.