
Introduction
Guanosine 3’, 5’-cyclic monophosphate (cGMP) is a key second messenger molecule in intracellular signal transduction. Monitoring cGMP levels is one of the most common ways to screen for agonists and antagonists of G protein–coupled receptors (GPCRs).
Assay type | Competitive |
Format | 96T |
Detection method | Colorimetric |
IC50 | 5 pmol/mL |
Detection limit | 0.1 pmol/mL |
Elabscience® cGMP ELISA Kit uses Competitive-ELISA as the method. The microtiter plate provided in this kit has been pre-coated with goat anti-mouse IgG. During the reaction, sample or standard, cGMP-HRP Conjugate and mouse monoclonal anti-cGMP antibody (cGMP MAb) are sequentially added to each wells. cGMP in the sample or standard competes with a fixed amount of HRP-labeled cGMP for the binding sites of cGMP MAb. Free components are washed from the plate. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop 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. The ELISA typically displays an IC50 (50% B/B0) of approximately 5 pmol/mL and a detection limit of approximately 0.1 pmol/mL.
Kit components
Item | Specification | Storage |
Goat anti-mouse IgG Micro-plate (12 × 8 wells) | 96 wells | 4℃ |
cGMP Standard (10000 pmol/mL) | 0.1 mL | -80℃ |
cGMP-HRP Conjugate (1000×) | 0.02 mL | -80℃ |
Anti-cGMP McAb (1000×) | 0.02 mL | -80℃ |
Assay Buffer (10×) | 30 mL | 4℃ |
Neutralizing Buffer | 8 mL | 4℃ |
Sample Diluent | 30 mL | 4℃ |
TMB Substrate | 18 mL | 4℃ |
Stop Solution | 8 mL | 4℃ |
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