Rabbit eNOS Polyclonal Antibody | anti-eNOS antibody

eNOS (Tyr-657)/nNOS (Tyr-895), phospho-specific Antibody

Cat. Num. AAA474477

• Gene Names: NOS3; eNOS; ECNOS
• Reactivity: Human, Rat, Mouse
• Applications: Western Blot, ELISA, Immunocytochemistry
• Synonyms: eNOS; Polyclonal Antibody; eNOS (Tyr-657)/nNOS (Tyr-895); phospho-specific Antibody; phospho-specific; NOSe 0657; anti-eNOS antibody

• Host: Rabbit
• Reactivity: Human, Rat, Mouse
• Clonality: Polyclonal
• Specificity: The antibody detects a 140kDa* band on SDS-PAGE immunoblots of humanuMbilical vein endothelial cells treated with pervanadate, and this reactivity is not observed after akaline phosphatase treatment.
• Form/Format: Rabbit polyclonal, affinity-purified antibody is supplied in 100ul phosphate-buffered saline, 50% glycerol, 1mg/ml BSA, and 0.05% sodium azide.

• Applicable Applications for anti-eNOS antibody: Western Blot (WB), ELISA (EIA), Immunocytochemistry (ICC)
• Application Notes: ELISA: 1:2000; ICC: 1:400; WB: 1:1000
• Immunogen: Phospho-eNOS (Tyr-657) synthetic peptide (coupled to carrier protein) corresponding to amino acids surrounding tyrosine 657 in human eNOS. This sequence is conserved in mouse (Tyr-656) and rat (Tyr-656) eNOS, and is identical to the conserved site in nNOS (Tyr-895). The site is also well conserved in iNOS (Tyr-631).
• Preparation and Storage: Store at -20 degree C. Stable for 1 year.

Western Blot (WB)

(Western blot analysis of humanuMbilical vein endothelial cells stimulated with pervanadate (1mM) for 30 min. (lanes 1, 3, & 5) then the blot was treated with alkaline phosphatase (lanes 2, 4, & 6). The blots were probed with anti-eNOS monoclonal antibody (NM2211; lanes 1 & 2), anti-eNOS (Tyr-657) phospho-specific antibody (NP4031; lanes 3 & 4), or anti-eNOS polyclonal antibody (NP2281; lanes 5 & 6).)

Immunocytochemistry (ICC)

(Immunocytochemical labeling of nNOS phosphorylation in rat PC12 cells differentiated with NGF. The cells were probed with mouse monoclonal (mAb) nNOS (NM4011), and rabbit polyclonal (pAb) nNOS (C-terminal region), nNOS (Tyr-895)/eNOS (Tyr-657), and nNOS (Tyr-1326)/iNOS (Tyr-1055). The antibodies were detected using appropriate secondary antibody conjugated to DyLight 594.)

Related Product Information for anti-eNOS antibody

Nitric oxide (NO) has a broad range of biological activitiesand is implicated in signaling pathways in phylogeneticallydiverse species. Nitric oxide synthases (NOS), theenzymes responsible for synthesis of NO, are homodimerswhose monomers are themselves two fused enzymes: acytochrome reductase and a cytochrome that requiresthree cosubstrates (L-arginine, NADPH, and oxygen) andfive cofactors or prosthetic groups (FAD, FMN, calmodulin, tetrahydrobiopterin, and heme). Several distinct NOSisoforms are produced from three distinct genes, inducibleNOS (iNOS, NOS-II), neuronal NOS (bNOS, NOS-I), andendothelial NOS (eNOS, ecNOS, NOS-III). Regulation ofeNOS activity occurs through phosphorylation at multiplesites. Phosphorylation of Ser-633 (mouse Ser-632) in theFMN binding domain increases eNOS activity and may beimportant for the maintenance of NO synthesis after initialactivation by Ca2+ flux and Ser-1177 phosphorylation. Tyr-657 is phosphorylated by PYK2 in response to fluid shearstress and this phosphorylation leads to attenuation ofenzyme activity.

References

Loot, A. E. et al. (2009) J Exp Med. 206(13):2889.
Fisslthaler, B. et al. (2008) Circ Res. 102:1520.
Mount, P.F. et al. (2007) J Mo.l Cell. Cardiol. 42(2):271.
Musicki, B. et al. (2005) Proc. Natl. Acad. Sci.102(33):11870.

NCBI and Uniprot Product Information

• NCBI GI #: 40254422
• NCBI GeneID: 4846
• NCBI Accession #: NP_000594.2
• NCBI GenBank Nucleotide #: NM_000603.4
• UniProt Accession #: P29474; Q13662; Q14251; Q14434; Q548C1; Q6GSL5; Q9UDC6; A0S0A7; A0S0A8; A8KA63; B2RCQ1; E9PFR2
• Molecular Weight: 133,289 Da
• NCBI Official Full Name: nitric oxide synthase, endothelial isoform 1
• NCBI Official Synonym Full Names: nitric oxide synthase 3 (endothelial cell)
• NCBI Official Symbol: NOS3
• NCBI Official Synonym Symbols: eNOS; ECNOS
• NCBI Protein Information: nitric oxide synthase, endothelial; cNOS; EC-NOS; NOSIII; NOS type III; endothelial NOS; constitutive NOS
• UniProt Protein Name: Nitric oxide synthase, endothelial
• UniProt Gene Name: NOS3
• UniProt Synonym Gene Names: cNOS; eNOS; NOSIII
• UniProt Entry Name: NOS3_HUMAN

NCBI Description

Nitric oxide is a reactive free radical which acts as a biologic mediator in several processes, including neurotransmission and antimicrobial and antitumoral activities. Nitric oxide is synthesized from L-arginine by nitric oxide synthases. Variations in this gene are associated with susceptibility to coronary spasm. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, May 2009]

Uniprot Description

eNOS: endothelial constitutive nitric oxide synthase. Synthesizes nitric oxide (NO) from arginine and oxygen, which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets. Induced in humans by caloric restriction. Stimulated by calcium/calmodulin. Inhibited by NOSIP and NOSTRIN.

Protein type: EC 1.14.13.39; Oxidoreductase; Amino Acid Metabolism - arginine and proline

Chromosomal Location of Human Ortholog: 7q36

Cellular Component: Golgi membrane; cytoskeleton; cytoplasm; plasma membrane; caveola; nucleus; cytosol

Molecular Function: calmodulin binding; actin monomer binding; protein binding; FAD binding; FMN binding; NADPH-hemoprotein reductase activity; nitric-oxide synthase activity; iron ion binding; heme binding; NADP binding; cadmium ion binding

Biological Process: regulation of systemic arterial blood pressure by endothelin; negative regulation of potassium ion transport; negative regulation of muscle hyperplasia; positive regulation of vasodilation; nitric oxide metabolic process; negative regulation of cell proliferation; negative regulation of hydrolase activity; regulation of blood pressure; regulation of the force of heart contraction by chemical signal; lipopolysaccharide-mediated signaling pathway; angiogenesis; negative regulation of blood pressure; regulation of blood vessel size; nitric oxide biosynthetic process; mitochondrion organization and biogenesis; interaction with host; negative regulation of calcium ion transport; in utero embryonic development; positive regulation of guanylate cyclase activity; regulation of sodium ion transport; ovulation from ovarian follicle; positive regulation of angiogenesis; response to heat; endothelial cell migration; arginine catabolic process; blood vessel remodeling; regulation of nitric-oxide synthase activity; vascular endothelial growth factor receptor signaling pathway; blood coagulation; nitric oxide mediated signal transduction; lung development; smooth muscle hyperplasia

Disease: Preeclampsia/eclampsia 1; Alzheimer Disease; Hypertension, Essential; Stroke, Ischemic