Catalog # NM2321

eNOS (Ser-632), phospho-specific Antibody

Mouse Monoclonal

Application / Dilution

ELISA	1:1000
WB	1:500

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 140 kDa

Isotype IgG1

Nitric oxide (NO) has a broad range of biological activities and is implicated in signaling pathways in phylogenetically diverse species. Nitric oxide synthases (NOS), the enzymes responsible for synthesis of NO, are homodimers whose monomers are themselves two fused enzymes: a cytochrome reductase and a cytochrome that requires three cosubstrates (L-arginine, NADPH, and oxygen) and five cofactors or prosthetic groups (FAD, FMN, calmodulin, tetrahydrobiopterin, and heme). Several distinct NOS isoforms are produced from three distinct genes. The inducible form of NOS, iNOS (NOS-II), is Ca2+ independent and is expressed in a broad range of cell types, and two constitutive Ca2+/CaM-dependent forms of NOS: nNOS (bNOS, NOS-I) identified in neurons and eNOS (ecNOS, NOS-III) identified in endothelial cells. Regulation of eNOS activity occurs through phosphorylation at multiple sites. Phosphorylation of Ser-633 (mouse Ser-632) in the FMN binding domain increases eNOS activity and may be important for the maintenance of NO synthesis after initial activation by Ca2+ flux and Ser-1177 phosphorylation.

References

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.
Xie, Q.W. et al. (1992) Science 256:225.

Western blot analysis of human umbilical vein endothelial cells stimulated with Calyculin A and untreated (lanes 1, 3, 5) or treated (lanes 2, 4, 6) with lambda phosphatase. The blots were probed with anti-endothelial Nitric Oxide Synthase (eNOS) monoclonal antibody (lanes 1 & 2), anti-eNOS (Ser-632) phospho-specific antibody (lanes 3 & 4), and anti-eNOS polyclonal antibody (lanes 5 & 6).

Clone M232 was generated from a Phospho-eNOS (Ser-632) synthetic peptide (coupled to carrier protein) corresponding to amino acids surrounding Ser-632 in mouse eNOS. This sequence is conserved in human (Ser-633) and rat (Ser-632) eNOS, and has low homology to other NOS family members.

*For more information, see UniProt Accession P70313

Mouse monoclonal antibody purified with protein A chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

The antibody detects a 140 and 120 kDa* bands on SDS-PAGE immunoblots of human umbilical vein endothelial cells, but these bands are not observed after lambda phosphatase treatment. The 120 kDa band may be a truncated form of eNOS.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: