Monoclonal Anti-cGMP EIA Kit without Acetylation

Product Highlight

• based on the unique mouse monoclonal Anti-cGMP antibody
  It is the only EIA kit on the market with monoclonal anti-cGMP antibody (all other kits are based on polyclonal anti-cGMP antibodies). 
   
• No more hazardous chemical (acetic anhydride ) for your cGMP assay 
  The monoclonal Anti-cGMP antibody has similar affinities for non-acetylated cGMP and acetylated cGMP. Thus acetylation with the acetic anhydride is permanently eliminated from the assay.
   
• Super sensitivity and selectivity 
  The monoclonal Anti-cGMP antibody displays >10⁸ fold of selectivity over cAMP, GTP, and other nucleoside analogues. 
  The Anti-cGMP EIA Kit provides significantly improved sensitivity and selectivity over other kits based on polyclonal anti-cGMP antibodies on the market.
   
• Shortest assay time and high-throughput format 
  The Anti-cGMP EIA Kit is the best choice for drug screenings.

Product Description

Guanosine 3',5'-cyclic monophosphate (cyclic GMP; cGMP) plays critical regulatory roles in many physiological processes. cGMP is produced from GTP by guanylyl cyclases and is degraded by phosphodiesterases. Stimulation of guanylyl cyclases or inhibition of phosphodiesterases can increase cellular cGMP concentrations. Inhibitors of the cGMP-specific phosphodiesterases are used for treating human diseases. For example, inhibitors of cGMP specific phosphodiesterase type 5 (such as Viagra and Cialis) are used for treating erectile dysfunction.

To screen for inhibitors of phosphodiesterases or stimulators of guanylyl cyclases, it is essential to have a fast, sensitive, selective and reproducible method to measure the cGMP concentrations. This is especially true for the initial screenings given the possible weaker effects of larger pools of compounds.

Currently available other ELISA kits measuring cGMP levels are based on the non-affinity-purified polyclonal anti-cGMP antibody. Despite the claimed selectivity, these polyclonal anti-cGMP antibodies display certain cross-reactivity with cAMP or GTP. In most cell types, cGMP is present at levels ~100 fold lower than cAMP.

NewEast Biosciences cGMP ELISA kit is based on the unique mouse monoclonal anti-cGMP antibody. This monoclonal anti-cGMP antibody displays >108 fold of selectivity over cAMP, GTP, and other nucleoside analogues. NewEast Biosciences cGMP ELISA kit provides significantly improved sensitivity and selectivity over other kits based on polyclonal anti-cGMP antibodies. Our monoclonal anti-cGMP antibody-based ELISA kit also avoids the batch-to-batch variations associated with polyclonal antibody productions from animals, thus providing the reproducibility in the long run.

Furthermore, while polyclonal anti-cGMP antibodies used in other ELISA kits have higher affinity for acetylated cGMP than non-acetylated cGMP, NewEast Biosciences monoclonal anti-cGMP antibody has similar affinities to non-acetylated and acetylated cGMP molecules. Therefore, acetylation treatments of samples and standards are not needed in NewEast Biosciences cGMP ELISA kit. This significantly reduces the time for the assay. The avoidance of organic reagents used in the acetylation process provides a safe and healthy work environment.

Principle Outline

NewEast Biosciences cGMP ELISA Kit is a competitive immunoassay to measure the cGMP levels, either from cell extracts or from in vitro guanylyl cyclase assays. Briefly, multi-well plates are coated with goat-anti-mouse serum. cGMP in cell extracts or in in vitro guanylyl cyclase assays will competitively bind to the monoclonal anti-cGMP antibody in the presence of fixed amounts of cGMP-conjugated horse-radish peroxidase. Known amounts of cGMP are used as standards to generate the calculation curve. After a short incubation, the excess reagents are washed away and substrate is added. The multiwell plates are then read on a microplate reader at 450 nm. The intensity of the yellow color is inversely proportional to the concentration of cGMP in samples. The measured optical density is used to calculate the concentration of cGMP in samples based on the curve from the cGMP standards.

Publications:

1.  Sulfhydryl-dependent dimerization of soluble guanylyl cyclase modulates the relaxation of porcine pulmonary arteries to nitric oxide     Pflugers Arch. 2013 Feb;465(2):333-41

2.  Pressure-overload-induced subcellular relocalization/oxidation of soluble guanylyl cyclase in the heart modulates enzyme stimulation     Circ Res. 2012 Jan 20;110(2):295-303

3.  Natriuretic peptides block synaptic transmission by activating phosphodiesterase 2A and reducing presynaptic PKA activity     Proc Natl Acad Sci U S A. 2012 Oct 23;109(43):17681-6

4.  Role of sulfhydryl-dependent dimerization of soluble guanylyl cyclase in relaxation of porcine coronary artery to nitric oxide     Cardiovasc Res. 2011 Jun 1;90(3):565-72

5.  Molt-inhibiting hormone from Chinese mitten crab (Eriocheir sinensis): Cloning, tissue expression and effects of recombinant peptide on ecdysteroid secretion of YOs     Gen Comp Endocrinol. 2011 Sep 15;173(3):467-74

6.  Inhibition of Striatal Soluble Guanylyl Cyclase-cGMP Signaling Reverses Basal Ganglia Dysfunction and Akinesia in Experimental Parkinsonism     PLoS One. 2011;6(11):e27187

7.  Molecular analysis, developmental function and heavy metal-induced expression of ABCC5 in zebrafish     Comp Biochem Physiol B Biochem Mol Biol. 2011 Jan;158(1):46-55

8.  Feedback control through cGMP-dependent protein kinase contributes to differential regulation and compartmentation of cGMP in rat cardiac myocytes     Circ Res. 2010 Nov 12;107(10):1232-40

9.  The Soluble Guanylate Cyclase Activator BAY 58-2667 Protects against Morbidity and Mortality in Endotoxic Shock by Recoupling Organ Systems     PLoS One. 2013 Aug 28;8(8):e72155

10.  Stimulation of guanylyl cyclase-D by bicarbonate     Biochemistry. 2009 May 26;48(20):4417-22

11.  Volume overload induces differential spatiotemporal regulation of myocardial soluble guanylyl cyclase in eccentric hypertrophy and heart failure     Journal of Molecular and Cellular Cardiology Volume 60, July 2013, Pages 72–83

12.  Defining Specificity Determinants of Cyclic GMP-Mediated Gustatory Sensory Transduction in Caenorhabditis elegans     genetics.113.152660v1 194/4/885

13.  The role of nitric oxide signaling in food intake; insights from the inner mitochondrial membrane peptidase 2 mutant mice     Redox Biology Volume 1, Issue 1, 2013, Pages 498–507

14.  Cross regulation between cGMP-dependent protein kinase and Akt in vasodilatation of porcine pulmonary artery     Journal of Cardiovascular Pharmacology: November 2014 - Volume 64 - Issue 5 - p 452-459

15.  Genetic ablation of solute carrier family 7a3a leads to hepatic steatosis in zebrafish during fasting     Hepatology Volume 60, Issue 6, pages 1929–1941, December 2014