HitHunter™ cGMP Assay

Guanosine 3'5'-cyclic monophosphate (cGMP) is an intracellular second messenger. Either receptor bound natriuretic peptides or nitrous oxide activate the production of cGMP from GTP. cGMP is a regulator of ion channel conductance, glycogenolysis, and cellular apoptosis. It also relaxes smooth muscle tissues. cGMP signaling is mediated by three effector protein:protein kinase G (PKG), phosphodiesterase 5 (PDE type 5) and cGMP regulated ion channels. HitHunter cGMP is a biochemical competitive immunoassay, based on β-galactosidase complementation.

Product Features

•Directly measure cGMP from cells in suspension
•HTS-friendly, chemiluminescent assay with high Z' (0.75)
•Scalable assay adaptable to 1536 applications
•Robust assays with large assay windows

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Applications

•Directly measure cGMP in cell-based assays
•Ideal for cGMP-dependent phosphodiesterases
•Determine activity of guanylate cyclase

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Available Collaterals

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Measuring cellular cGMP with EFC chemiluminescent detection

HitHunter cGMP is an in-vitro based competitive immunoassay. Free cGMP from cell lysates compete for antibody binding against labeled ED-cGMP conjugate, a small peptide fragment of β-galactosidase (β-gal). Unbound ED-cGMP is free to complement with EA, an inactive β-gal EFC enzyme, to form active enzyme, which subsequently hodrolyzes luminescent substrate. The amount of luminescent signal produced is directly proportional to the amount of free cGMP in the cell lysate.