Our eNOS (Tyr-657)/nNOS (Tyr-895) rabbit polyclonal phosphospecific primary antibody from PhosphoSolutions is produced in-house. It detects human, mouse, and rat eNOS (Tyr-657)/nNOS (Tyr-895) and is antigen affinity purified. It is great for use in WB, ICC.
Western blot analysis of human umbilical vein endothelial cells stimulated with pervanadate (1 mM) 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).
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.
Antigen Affinity Purified
Polyclonal
IgG
ICC, WB
Rabbit
NOS3
140
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).
Human, Mouse, Rat
Storage at -20°C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20°C.
Liquid
PBS + 1 mg/ml BSA, 0.05% NaN3 and 50% glycerol
WB: 1:1000
ICC: 1:400
Unconjugated
The antibody detects a 140 kDa* band on SDS-PAGE immunoblots of human umbilical vein endothelial cells treated with pervanadate, and this reactivity is not observed after akaline phosphatase treatment.
Phosphorylated
Tyr-657
Western blots performed on each lot.
For research use only. Not intended for therapeutic or diagnostic use. Use of all products is subject to our terms and conditions, which can be viewed on our website.
United States
After date of receipt, stable for at least 1 year at -20°C.
