eNOS (Endothelial Nitric Oxide Synthase, NOS 3, NOS III)

Nitric oxide (NO) is a potent bio-regulatory molecule, which plays important roles in the regulation of a variety of normal developmental and physiological processes (1,3,4). NO is generated by nitric oxide synthase (NOS) in a reaction which converts L-arginine and oxygen into citrulline and nitric oxide (1,3,4). Three distinct mammalian genes have been identified which encode NOS isoforms: neuronal (nNOS), macrophage or inducible (iNOS) and endothelial (eNOS) (1-4). The NOS isoforms can be subdivided into two general categories, constitutive or inducible, based on differences in their regulation and activities (1). The constitutive isoforms include eNOS and nNOS (1,3,4). These isoforms are always present but remain inactive until intracellular calcium levels increase resulting in enhanced calcium/calmodulin binding and subsequent activation (1,3,4). In contrast, the inducible NOS isoform, iNOS, is not normally expressed, but is induced (though alterations in gene expression, mRNA stability or protein synthesis) in response to certain cytokines (1,3,4). The nomenclature of the NOS family members suggests restricted isoform expression patterns; however, this turns out to be somewhat misleading. For example, eNOS expression is not restricted to the endothelium. It has been detected in the endothelium of blood vessels, the epithelium of some tissues including the bronchial tree, and in neurons of the brain especially in the pyramidal cells of the hippocampus. In addition, iNOS not only occurs in macrophages but has been detected in several other cell types including: hepatocytes, chrondrocytes, endothelial cells and fibroblasts. eNOS is a ~135kD protein and is the only NOS isoform known to contain an N-terminal myristoylation site (1-7). Myristoylation serves to target the eNOS protein to intracellular membranes thereby restricting NO signaling to specific subcellular compartments (1-4). The eNOS protein shares the highest degree of identity with the nNOS protein (1-4). eNOS activity accounts for endothelium-dependent blood vessel relaxation and has, therefore, been implicated in the blood pressure regulation (1,2,7). This was role of eNOS was confirmed in eNOS knockout mice which exhibited a 35% higher mean blood pressure than control animals (7). However, this study also found that the eNOS protein is not responsible for maintenance of blood pressure (7). \n\nApplications: \nSuitable for use in ELISA, Western Blot and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution:\nELISA: 0.1-1ug/ml \nWestern Blot: 1-2ug/ml\nImmunoprecipitation: 5ug/IP \nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts

SPECIFICATIONS

Catalog Number

E3010-84

Size

100ug

Applications

ELISA, IP, WB

Hosts

Mouse

Reactivities

Bov

Form

Supplied as a liquid in PBS, pH 7.4, 0.1% sodium azide.

P Type

Mab

Purity

Purified by Protein A affinity chromatography.

Isotype

IgG1,k

References

1. Sessa, W.C., J. Vasc. Res. (1994) 31:131-143. 2. Snyder, S., (1995) Nature 377:196-197. 3. Lowenstein, C.L., et al., (1994) Annals of Int. Med. 120:227-237. 4. Nathan, C., et al., (1994) J. Biol. Chem. 269:13,725-13,728. 5. Sessa, W.C., et al., (1992) J. Biol. Chem. 267:15,274-15,276. 6. Sessa, W.C., et al., (1995) J. Biol. Chem. 270:17,641-17,644. 7. Huang, P.L., et al., (1995) Nature 377:239-242.

Additional Info

Specific for endothelial nitric oxide synthase (eNOS) and does not crossreact with either inducible iNOS or neuronal nNOS. The immunoreactive epitope has not been localized, however, this antibody recognizes an epitope distinct from that recognized by Cat. #E3010-86. Species Crossreactivity: Bovine, likely to react with other mammalian species including mouse and human eNOS proteins based on sequence homology. Lysates Tested: Bovine aortic endothelial cells, and 293 cells stably expressing recombinant bovine eNOS.