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Protocol
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This ELISA Kit uses Competitive-ELISA as the method. The microtiter plate provided in this kit has been pre-coated with cGMP. During the reaction, cGMP in the sample or standard competes with a fixed amount of cGMP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to cGMP. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of cGMP in the samples is then determined by comparing the OD of the samples to the standard curve. 6th Edition, revised in June, 2015 www.elabscience.com 4
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