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This assay employs the quantitative sandwich enzyme immunoassay technique. Anantibody specific for human VEGF has been pre-coated onto a microplate. Standardsand samples are pipetted into the wells and any human VEGF present is bound by theimmobilized antibody. After washing away any unbound substances, an enzyme-linkedantibody specific for human VEGF is added to the wells. Following a wash to removeany unbound antibody-enzyme reagent, TMB substrate (Chromogenic agent) is addedto the wells and color develops in proportion to the amount of human VEGF boundinthe initial step. The color development is stopped and the intensity of the color ismeasured.