cAMP Gs dynamic kit
Optimal performance when applied to Gs activation or inhibition
The cGMP assay kit accurately measures Cyclic GMP produced by cells. The assay can be used to screen molecules that modulate the Phosphodiesterase (PDE) activity. cGMP is a major second messenger that mediates cell activities by activating various protein effectors. Cellular cGMP is synthesized by soluble guanylyl cyclases and particulate guanylyl cyclase, triggered by nitric oxide and atrial natriuretic peptide (ANP) binding respectively. The cGMP kit offers superior benefits over traditional ELISA readouts with the ease-of-use and high sensitivity of homogeneous TR-FRET assays.
The cGMP assay features a streamlined protocol with only two incubation steps:
- Cell stimulation by target compounds
- cGMP detection using HTRF reagents.
This protocol requires only a single, one-hour incubation period following cell stimulation.
|Working range:||0.5 to 500 nM|
|Detection limit:||0.7 nM|
|Specificity:||cGMP 100%, GMP <0.003%, |
AMP, ATP &cAMP <0.001%
|Gene family||Major tissue expression||Substrate|
|PDE2||Adrenal cortex, brain, heart||cAMP/cGMP|
|PDE3||Heart, adipose tissue, pancreas, platelets||cAMP/cGMP|
|PDE5||Lung, platelets, smooth muscle, corpus cavernosum||cGMP|
|PDE6||Rod and cone photoreceptor outer segments||cGMP|
|PDE7||Skeletal muscle, T-cells, B-cells||cAMP|
|PDE8||Testis, liver, thyroid||cGMP|
|PDE11||Skeletal muscle, prostate||cAMP/cGMP|
Increases in IW-1973 increases both cGMP production and VASP phosphorylation. IW-1973 is a sGC stimulator. cGMP production and VASP phosphorylation were measured using the HTRF assays.
Adapted from Tobin J.V, et al (2018). Pharmacological characterization of IW-1973, a novel soluble guanylate cyclase stimulator with extensive tissue distribution, anti-hypertensive, anti- inflammatory,and anti-fibrotic effects in preclinical models of disease. J Pharmacol Exp Ther 365(3):664-675
Nitric Oxide is a key effector protein that - among other roles - is involved in cardiovascular hemostasis and vascular relaxation. In the event of a vascular stress, NO is synthesized at its location. It binds to sGC on the environing tissues, triggering the downstream decrease of Ca2+ and increase of cGMP, which in turn alters the activity of a collection of protein kinase G, cyclic nucleotide-gated ion channels and phosphodiesterases, leading to the vasodilatation of nearby vessels, blood flow increase and lowered tension.
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