Assay Principle: 

The DetectX® Cyclic GMP Direct ELISA Kit – Improved Sensitivity quantitatively measures Cyclic GMP (cGMP) in saliva, urine, plasma, cell and tissue lysates, and tissue culture media. This competitive ELISA has a run time of 2.5 hours. Please read the complete kit insert for more information before performing this assay. Use our provided cGMP standard to generate a standard curve for the assay.

Protocol Summary:

• Introduce standards or diluted samples into the provided transparent microtiter plate, pre-coated with goat anti-rabbit IgG antibody.
• Add cGMP peroxidase conjugate and cGMP rabbit polyclonal antibody to initiate the immunological reaction.
• Incubate the mixture covered at room temperature with shaking for 2 hours. The reaction varies inversely with the cGMP concentration in the sample.
• After incubation, remove excess conjugate and add the TMB substrate, which reacts with the bound conjugate, producing a measurable signal.
• Use a plate reader to detect the signal at 450nm and calculate the cGMP concentration based on the standard curve.

Background:

Guanosine 3’, 5’-cyclic monophosphate (cyclic GMP; cGMP) is a n important second messenger in various biological processes. cGMP circulates at 10-100 times lower concentrations than cAMP in most tissues. It exerts its regulatory functions through cGMP-dependent kinases, ion channels or transporters, and phosphodiesterase-mediated degradation. Guanylate cyclases, which catalyze the conversion of GTP to cGMP, are either soluble or membrane-bound. Soluble guanylate cyclases are responsive to nitric oxide, while membrane-bound guanylate cyclases respond to hormones like acetylcholine, insulin, and oxytocin. Other agents, including serotonin and histamine, can also elevate cGMP levels. Understanding cGMP dynamics is vital in research areas focusing on cardiovascular and nervous system functions, as well as in studying cellular signal transduction pathways.

The DetectX® Cyclic GMP Direct ELISA Kit is an essential tool for researchers studying cellular composition, signal transduction, and various physiological processes regulated by cGMP. This kit’s improved sensitivity and specificity make it ideal for accurate cGMP quantification in diverse sample types.