Product: Phospho-eNOS (Ser1177) Antibody
Catalog: AF3247
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Rabbit, Dog
Mol.Wt.: 140kD; 133kD(Calculated).
Uniprot: P29474
RRID: AB_2834673

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:200, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(82%), Rabbit(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
Phospho-eNOS (Ser1177) Antibody detects endogenous levels of eNOS only when phosphorylated at Serine 1177.
RRID:
AB_2834673
Cite Format: Affinity Biosciences Cat# AF3247, RRID:AB_2834673.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

cNOS; Constitutive NOS; EC NOS; EC-NOS; ecNOS; Endothelial nitric oxidase synthase; Endothelial nitric oxide synthase; Endothelial nitric oxide synthase 3; Endothelial NOS; eNOS; Nitric oxide synthase 3 (endothelial cell); Nitric oxide synthase 3; Nitric oxide synthase 3 endothelial cell; Nitric oxide synthase endothelial; Nitric oxide synthase, endothelial; NOS 3; NOS III; NOS type III; NOS3; NOS3_HUMAN; NOSIII;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P29474 NOS3_HUMAN:

Platelets, placenta, liver and kidney.

Description:
eNOS is an 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.
Sequence:
MGNLKSVAQEPGPPCGLGLGLGLGLCGKQGPATPAPEPSRAPASLLPPAPEHSPPSSPLTQPPEGPKFPRVKNWEVGSITYDTLSAQAQQDGPCTPRRCLGSLVFPRKLQGRPSPGPPAPEQLLSQARDFINQYYSSIKRSGSQAHEQRLQEVEAEVAATGTYQLRESELVFGAKQAWRNAPRCVGRIQWGKLQVFDARDCRSAQEMFTYICNHIKYATNRGNLRSAITVFPQRCPGRGDFRIWNSQLVRYAGYRQQDGSVRGDPANVEITELCIQHGWTPGNGRFDVLPLLLQAPDDPPELFLLPPELVLEVPLEHPTLEWFAALGLRWYALPAVSNMLLEIGGLEFPAAPFSGWYMSTEIGTRNLCDPHRYNILEDVAVCMDLDTRTTSSLWKDKAAVEINVAVLHSYQLAKVTIVDHHAATASFMKHLENEQKARGGCPADWAWIVPPISGSLTPVFHQEMVNYFLSPAFRYQPDPWKGSAAKGTGITRKKTFKEVANAVKISASLMGTVMAKRVKATILYGSETGRAQSYAQQLGRLFRKAFDPRVLCMDEYDVVSLEHETLVLVVTSTFGNGDPPENGESFAAALMEMSGPYNSSPRPEQHKSYKIRFNSISCSDPLVSSWRRKRKESSNTDSAGALGTLRFCVFGLGSRAYPHFCAFARAVDTRLEELGGERLLQLGQGDELCGQEEAFRGWAQAAFQAACETFCVGEDAKAAARDIFSPKRSWKRQRYRLSAQAEGLQLLPGLIHVHRRKMFQATIRSVENLQSSKSTRATILVRLDTGGQEGLQYQPGDHIGVCPPNRPGLVEALLSRVEDPPAPTEPVAVEQLEKGSPGGPPPGWVRDPRLPPCTLRQALTFFLDITSPPSPQLLRLLSTLAEEPREQQELEALSQDPRRYEEWKWFRCPTLLEVLEQFPSVALPAPLLLTQLPLLQPRYYSVSSAPSTHPGEIHLTVAVLAYRTQDGLGPLHYGVCSTWLSQLKPGDPVPCFIRGAPSFRLPPDPSLPCILVGPGTGIAPFRGFWQERLHDIESKGLQPTPMTLVFGCRCSQLDHLYRDEVQNAQQRGVFGRVLTAFSREPDNPKTYVQDILRTELAAEVHRVLCLERGHMFVCGDVTMATNVLQTVQRILATEGDMELDEAGDVIGVLRDQQRYHEDIFGLTLRTQEVTSRIRTQSFSLQERQLRGAVPWAFDPPGSDTNSP

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Dog
100
Rabbit
100
Horse
82
Sheep
0
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P29474 As Substrate

Site PTM Type Enzyme
K5 Methylation
T33 Phosphorylation
S53 Phosphorylation
Y81 Phosphorylation
C94 S-Nitrosylation
C99 S-Nitrosylation
S102 Phosphorylation
S114 Phosphorylation Q00535 (CDK5)
C184 S-Nitrosylation
C201 S-Nitrosylation
Y210 Phosphorylation
C212 S-Nitrosylation
S260 Phosphorylation
T495 Phosphorylation Q04759 (PRKCQ) , Q13237 (PRKG2) , P05771 (PRKCB) , P17612 (PRKACA) , Q02156 (PRKCE) , O75116 (ROCK2) , P17252 (PRKCA) , P05129 (PRKCG) , P24723 (PRKCH) , Q05655 (PRKCD) , Q13131 (PRKAA1) , P41743 (PRKCI) , Q05513 (PRKCZ)
S508 Phosphorylation
T512 Phosphorylation
S615 Phosphorylation P17612 (PRKACA) , Q9Y243 (AKT3) , P31751 (AKT2) , P31749 (AKT1)
S633 Phosphorylation Q13237 (PRKG2) , P54646 (PRKAA2) , Q13131 (PRKAA1) , P17612 (PRKACA)
S634 Phosphorylation
T636 Phosphorylation
S638 Phosphorylation
Y657 Phosphorylation Q14289 (PTK2B)
C661 S-Nitrosylation
S738 Phosphorylation
K773 Ubiquitination
C802 S-Nitrosylation
K834 Ubiquitination
S836 Phosphorylation
C853 S-Nitrosylation
T854 Phosphorylation
S870 Phosphorylation
K904 Ubiquitination
C976 S-Nitrosylation
C991 S-Nitrosylation
K1035 Ubiquitination
C1048 S-Nitrosylation
C1050 S-Nitrosylation
K1085 Ubiquitination
T1094 Phosphorylation
C1114 S-Nitrosylation
T1175 Phosphorylation
S1177 O-Glycosylation
S1177 Phosphorylation P22612 (PRKACG) , Q9Y478 (PRKAB1) , P31749 (AKT1) , Q13131 (PRKAA1) , P54646 (PRKAA2) , Q13237 (PRKG2) , P17612 (PRKACA)
S1179 Phosphorylation P31749 (AKT1)

Research Backgrounds

Function:

Produces nitric oxide (NO) 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.

Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.

PTMs:

Phosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity (By similarity). Phosphorylation of Ser-114 by CDK5 reduces activity.

Subcellular Location:

Cell membrane. Membrane>Caveola. Cytoplasm>Cytoskeleton. Golgi apparatus.
Note: Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle; which is favored by interaction with NOSIP and results in a reduced enzymatic activity.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Platelets, placenta, liver and kidney.

Subunit Structure:

Homodimer. Interacts with NOSIP and NOSTRIN. Interacts with HSP90AB1.

Family&Domains:

Belongs to the NOS family.

Research Fields

· Environmental Information Processing > Signal transduction > Calcium signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Apelin signaling pathway.   (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Metabolism > Amino acid metabolism > Arginine biosynthesis.

· Metabolism > Amino acid metabolism > Arginine and proline metabolism.

· Metabolism > Global and overview maps > Metabolic pathways.

· Organismal Systems > Immune system > Platelet activation.   (View pathway)

· Organismal Systems > Endocrine system > Estrogen signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

References

1). Guo J et al. Endothelial SIRT6 Is Vital to Prevent Hypertension and Associated Cardiorenal Injury Through Targeting Nkx3.2-GATA5 Signaling. Circ Res 2019 May 10;124(10):1448-1461 (PubMed: 30894089) [IF=14.467]

2). Liao L et al. Leonurine Ameliorates Oxidative Stress and Insufficient Angiogenesis by Regulating the PI3K/Akt-eNOS Signaling Pathway in H2O2-Induced HUVECs. Oxid Med Cell Longev 2021 Aug 3;2021:9919466. (PubMed: 34394836) [IF=5.076]

3). Cai BB et al. Acid sphingomyelinase downregulation alleviates vascular endothelial leptin resistance in rats. Acta Pharmacol Sin 2019 Dec 17 (PubMed: 31848475) [IF=5.064]

Application: WB    Species: Rat    Sample: rat aortic endothelial cells (RAECs)

Fig. 4 ASM downregulation increased eNOS/NO signaling in RAECs. RAECs were incubated with 0.3 mM PA for 24 h and then transfected with ASM siRNA, or RAECs were transfected with Ob-Rb siRNA for 48 h and then treated with 10 μM AMI and 10 μM IMI for 24 h. Then, 100 nM leptin was added for 15 min prior to the collection of the cells. Summarized data showing the effects of ASM downregulation on the ratio of p-eNOS (Ser1177)/eNOS (a), eNOS activity (b), NO release (c), Ob-Rb expression (d), and the ratios of p-STAT3/STAT3 (e) and p-Akt/Akt (f) in RAECs. *P < 0.05 vs. vehicle control (Vehl Scr); #P < 0.05 vs. the PA alone-treated group; ΔP < 0.05 vs. the Ob-Rb siRNA alone-transfected group. Representative Western blot gels and summarized data showing the ratios of p-STAT3/STAT3 and p-Akt/Akt in RAECs that were pretreated with 10 μM cryptotanshinone for 24 h and then incubated with 0.3 mM PA for 24 h (g, h) or transfected with Ob-Rb siRNA for 48 h (i, j). *P < 0.05 vs. vehicle control (Vehl Scr); #P < 0.05 vs. the SCH772984 alone-treated group. The co-immunoprecipitation assay was used to test the interaction between p-STAT3 and p-Akt in RAECs treated with PA (k) or transfected with Ob-Rb siRNA (l). The data are the means ± SEMs from three experiments.

4). Xue M et al. Corin plays a protective role via upregulating MAPK and downregulating eNOS in diabetic nephropathy endothelial dysfunction. FASEB J 2020 Jan;34(1):95-106. (PubMed: 31914697) [IF=4.966]

Application: WB    Species: Human    Sample: HK‐2 cells

FIGURE 4 The effect of Corin deficiency in HK‐2 cells on EA.hy926 cells endothelial function and the mechanism. A, The wound healing scratch assay showed the effect of culture supernatants of HK‐2 cells on EA.hy926 cells migration at 0 and 24 h in Ctrl, NC‐siRNA, Corin‐siRNA, and Corin‐siRNA + ANP groups (original magnification 100×). B‐E, The ERK, P38, JNK, and their phosphorylation forms protein levels of EA.hy926 cells treated with culture supernatants of HK‐2 cells were measured by western blot (n = 6 for each group). F,G, The eNOS and p‐eNOS protein levels of EA.hy926 cells treated with culture supernatants of HK‐2 cells were measured by western blot (n = 6 for each group). Data are presented as mean ± SEM. *P < .05 and **P < .01

5). Lu Y et al. Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and palmitic acid. Biomed Pharmacother 2018 Nov;107:1631-1640 (PubMed: 30257381) [IF=4.545]

6). Gao Y et al. Ginsenoside Re inhibits PDGF-BB-induced VSMC proliferation via the eNOS/NO/cGMP pathway. Biomed Pharmacother 2019 May 10;115:108934 (PubMed: 31082773) [IF=4.545]

7). Yang R et al. Cyclocarya paliurus triterpenoids attenuate glomerular endothelial injury in the diabetic rats via ROCK pathway. J Ethnopharmacol 2022 Feb 24;291:115127. (PubMed: 35219820) [IF=3.690]

8). Qu K et al. Buxuhuayu decoction accelerates angiogenesis by activating the PI3K-Akt-eNOS signalling pathway in a streptozotocin-induced diabetic ulcer rat model . J Ethnopharmacol 2021 Jun 12;273:113824. (PubMed: 33581257) [IF=3.690]

9). Zhang W et al. Role of EGFR/ErbB2 and PI3K/AKT/e-NOS in Lycium barbarum polysaccharides Ameliorating Endothelial Dysfunction Induced by Oxidative Stress. Am J Chin Med 2019 Oct 23:1-17 (PubMed: 31645123) [IF=3.682]

10). Guo Y et al. Ketogenic diet aggravates hypertension via NF-κB-mediated endothelial dysfunction in spontaneously hypertensive rats. Life Sci 2020 Jul 21;258:118124. (PubMed: 32702443) [IF=3.647]

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