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). Suitable for use in ELISA, Western Blot and Immunoprecipitation. Other applications not tested.

Optimal dilutions to be determined by the researcher.

May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Purified bovine recombinant eNOS protein

Supplied as a liquid in PBS, pH 7.4, 0.09% sodium azide.

Purified by Protein A affinity chromatography.

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.