Product Name:
NOS3-pT1175+pS1177
Product Number:
ab-pn712
Target Full Name: Nitric oxide synthase, endothelial
Target Alias: CNOS; Constitutive NOS; ECNOS; EC-NOS; Endothelial nitric oxide synthase; Endothelial NOS; Nitric oxide synthase 3 (endothelial cell); Nitric-oxide synthase, endothelial; NOS type III;NOSIII;Endothelial NOS;eNOS;Constitutive NOS;cNOS; NOS, type III; NOS3; NOSIII
Product Type Specific: NOS3 phosphosite-specific antibody
Antibody Code: PN712
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T1175+S1177
Protein UniProt: P29474
Protein SigNET: NOS3
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human NOS3
Antibody Immunogen Sequence: RIR(pT)Q(pS)FSL(βA)C
Antibody Immunogen Description: Corresponds to amino acid residues R1172 to L1180. T1175+S1177 phosphorylation stimulates enzyme activity. These are two of the major in vivo phosphorylation sites in NOS3 (≥17 and >22 reports, respectively, by mass spectrometry). These phosphosites are not highly conserved in vertebrates, but are found in the rat. NOS3 is known to be phosphorylated in vitro at S1177 by Akt1 (PKBa), AMPKa1 (PRKAA1), PKACa (PRKACA), PKCd (PRKCD), and PKG2 (PRKG2).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.75 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Rat | Mouse.
Related Product 1: NOS2 S745 phosphorylation antibody (Cat. No.: AB-PN711)
Related Product 2: NOS1 S746 phosphorylation antibody (Cat. No.: AB-PN762)
Related Product 3: NOS2 Y151 phosphorylation antibody (Cat. No.: AB-PN763)
Scientific Background: NOS3 (Nitric oxide synthase, endothelial; eNOS) is a critical, constitutively expressed enzyme responsible for producing nitric oxide (NO) from L-arginine in the vascular endothelium. It catalyzes the reaction: 2 L-arginine + 3 NADPH + 4 O2 + H+ = 2 L-citrulline + 2 nitric oxide + 3 NADP+ + 4 H2O. It is essential for maintaining cardiovascular health by regulating vascular tone, promoting angiogenesis, and preventing platelet aggregation and leukocyte adhesion. NO diffuses to vascular smooth muscle cells, stimulating cGMP-mediated relaxation (vasodilation). It is a 133 kDa homodimeric enzyme composed of an N-terminal oxygenase domain and a C-terminal reductase domain. Its activity is calcium-dependent and regulated by phosphorylation (notably at Ser-1177 and Thr-495) and binding with caveolin-1, which inhibits the enzyme until activated by calcium/calmodulin. It requires as cofactors Heme b, FAD, FMN, calmodulin, and tetrahydrobiopterin (BH4) for proper function. It is primarily located in the Golgi apparatus and caveolae (small membrane invaginations) of endothelial cells, and distributed in platelets, placenta, liver and kidney. Isoform eNOS13C lacks eNOS activity, and is dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1. When cofactors like BH4 are deficient, the enzyme becomes "uncoupled" and produces superoxide (O2-) instead of NO, and contributes to cardiovascular disease. Variations in the NOS3 gene (such as Glu298Asp) are associated with reduced NO production, increased hypertension risk, and coronary artery disease. Reduced activity of NOS3 is linked to atherosclerosis, hypertension, and diabetes, while higher expression is sometimes associated with poor prognosis in certain cancers (e.g., gastric cancer). This description may include information annotated by UniProt and/or Google AI.