Principle of Assay
cGMP ELISA Kit employs the Competitive Enzyme Immunoassay (ELISA) technique for the quantitative measurement ofuniversalcGMP in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. A monoclonal antibody specific for cGMP has been pre-coated onto a 96-well microtiter plate. During the incubation, cGMP in the sample or standard competes with a fixed amount of Biotinylated-cGMP. Following incubation, Biotinylated-cGMP and free cGMP in the sample or standard are washed from the plate. Avidin conjugated to Horseradish Peroxidase (Avidin-HRP) is then added to each well and incubated. TMB substrate solution is then added to each well, and after incubation, the enzyme-substrate reaction is terminated by the addition of sulfuric acid stop solution. The color change is measured spectrophotometrically at a wavelength of 450nm, and the concentration of cGMP in the samples is then determined by comparing the O.D. 450nm of the samples to the standard curve. The O.D. 450nm is inversely proportional to the concentration of cGMP in the sample.
