Phospho-eNOS (Ser1177) Antibody - #AF3247

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产品: 磷酸化 eNOS (Ser1177) 抗体
货号: AF3247
描述: Rabbit polyclonal antibody to Phospho-eNOS (Ser1177)
应用: WB IHC IF/ICC
反应: Human, Mouse, Rat
预测: Pig, Bovine, Horse, Rabbit, Dog
分子量: 130~160kD; 133kD(Calculated).
蛋白号: P29474
RRID: AB_2834673

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说明书
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实验方案
WB Handbook
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IF/ICC Protocol

产品描述

来源:
Rabbit
应用:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡(IHC-p)或冰冻(IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA(peptide): 适用于抗原肽的ELISA检测.

反应:
Human,Mouse,Rat
预测:
Pig(100%), Bovine(100%), Horse(82%), Rabbit(100%), Dog(100%)
克隆:
Polyclonal
特异性:
Phospho-eNOS (Ser1177) Antibody detects endogenous levels of eNOS only when phosphorylated at Serine 1177.
RRID:
AB_2834673
引用格式: Affinity Biosciences Cat# AF3247, RRID:AB_2834673.
偶联:
Unconjugated.
纯化:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
保存:
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.
别名:

展开/折叠

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;

抗原和靶标

免疫原:
Uniprot:

P29474(NOS3_HUMAN) >>Visit HPA database.

基因/基因ID:
NOS3(4846)
表达:
P29474 NOS3_HUMAN:

Platelets, placenta, liver and kidney.

描述:
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.
序列:
MGNLKSVAQEPGPPCGLGLGLGLGLCGKQGPATPAPEPSRAPASLLPPAPEHSPPSSPLTQPPEGPKFPRVKNWEVGSITYDTLSAQAQQDGPCTPRRCLGSLVFPRKLQGRPSPGPPAPEQLLSQARDFINQYYSSIKRSGSQAHEQRLQEVEAEVAATGTYQLRESELVFGAKQAWRNAPRCVGRIQWGKLQVFDARDCRSAQEMFTYICNHIKYATNRGNLRSAITVFPQRCPGRGDFRIWNSQLVRYAGYRQQDGSVRGDPANVEITELCIQHGWTPGNGRFDVLPLLLQAPDDPPELFLLPPELVLEVPLEHPTLEWFAALGLRWYALPAVSNMLLEIGGLEFPAAPFSGWYMSTEIGTRNLCDPHRYNILEDVAVCMDLDTRTTSSLWKDKAAVEINVAVLHSYQLAKVTIVDHHAATASFMKHLENEQKARGGCPADWAWIVPPISGSLTPVFHQEMVNYFLSPAFRYQPDPWKGSAAKGTGITRKKTFKEVANAVKISASLMGTVMAKRVKATILYGSETGRAQSYAQQLGRLFRKAFDPRVLCMDEYDVVSLEHETLVLVVTSTFGNGDPPENGESFAAALMEMSGPYNSSPRPEQHKSYKIRFNSISCSDPLVSSWRRKRKESSNTDSAGALGTLRFCVFGLGSRAYPHFCAFARAVDTRLEELGGERLLQLGQGDELCGQEEAFRGWAQAAFQAACETFCVGEDAKAAARDIFSPKRSWKRQRYRLSAQAEGLQLLPGLIHVHRRKMFQATIRSVENLQSSKSTRATILVRLDTGGQEGLQYQPGDHIGVCPPNRPGLVEALLSRVEDPPAPTEPVAVEQLEKGSPGGPPPGWVRDPRLPPCTLRQALTFFLDITSPPSPQLLRLLSTLAEEPREQQELEALSQDPRRYEEWKWFRCPTLLEVLEQFPSVALPAPLLLTQLPLLQPRYYSVSSAPSTHPGEIHLTVAVLAYRTQDGLGPLHYGVCSTWLSQLKPGDPVPCFIRGAPSFRLPPDPSLPCILVGPGTGIAPFRGFWQERLHDIESKGLQPTPMTLVFGCRCSQLDHLYRDEVQNAQQRGVFGRVLTAFSREPDNPKTYVQDILRTELAAEVHRVLCLERGHMFVCGDVTMATNVLQTVQRILATEGDMELDEAGDVIGVLRDQQRYHEDIFGLTLRTQEVTSRIRTQSFSLQERQLRGAVPWAFDPPGSDTNSP

种属预测

种属预测:

score>80的预测可信度较高，可尝试用于WB检测。*预测模型主要基于免疫原序列比对，结果仅作参考，不作为质保凭据。

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

翻译修饰 - P29474 作为底物

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)

研究背景

功能:

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.

翻译修饰:

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.

细胞定位:

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
组织特异性:

Platelets, placenta, liver and kidney.

亚基结构:

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

蛋白家族:

Belongs to the NOS family.

研究领域

· 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.

文献引用

1). Endothelial SIRT6 Is Vital to Prevent Hypertension and Associated Cardiorenal Injury Through Targeting Nkx3.2-GATA5 Signaling. CIRCULATION RESEARCH, 2019 (PubMed: 30894089) [IF=20.1]
2). Ethanolic extract of Cissus quadrangularis improves vasoreactivity by modulation of eNOS expression and oxidative stress in spontaneously hypertensive rats. CLINICAL AND EXPERIMENTAL HYPERTENSION, 2022 (PubMed: 34648416) [IF=12.3]
3). Acid sphingomyelinase downregulation alleviates vascular endothelial leptin resistance in rats. ACTA PHARMACOLOGICA SINICA, 2020 (PubMed: 31848475) [IF=8.2]

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). Ginsenoside Re inhibits PDGF-BB-induced VSMC proliferation via the eNOS/NO/cGMP pathway. BIOMEDICINE & PHARMACOTHERAPY, 2019 (PubMed: 31082773) [IF=7.5]
5). Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and palmitic acid. BIOMEDICINE & PHARMACOTHERAPY, 2018 (PubMed: 30257381) [IF=7.5]
6). Ketogenic diet aggravates hypertension via NF-κB-mediated endothelial dysfunction in spontaneously hypertensive rats. LIFE SCIENCES, 2020 (PubMed: 32702443) [IF=6.1]

Application: WB    Species: Human    Sample: HUVECs

Fig. 5. Ketone body reduced eNOS expression via NF-κB in human umbilical vein endothelial cells. (A) eNOS, CD31 and α-SMA expression in HUVECs subjected to TGF-β and BHB for 48 h were measured with western blot. The quantified eNOS, CD31 and α-SMA levels are presented in (B-D). (E-I) The ERK and NF-κB signaling pathway and related inflammatory cytokine were measured with western blot. (J) Inhibition of NFκB prevented the effect of ketone body and TGF-β on eNOS expression in HUVECs. (K-M) Quantification of eNOS, CD31 and α-SMA. All data are expressed as a mean ± SEM. n = 6 per group. *P < 0.05; **P < 0.01; BHB, β-hydroxybutyrat; HUVECs, human umbilical vein endothelial cells; TGF-β, transforming growth factor-β.
7). Role of EGFR/ErbB2 and PI3K/AKT/e-NOS in Lycium barbarum polysaccharides Ameliorating Endothelial Dysfunction Induced by Oxidative Stress. AMERICAN JOURNAL OF CHINESE MEDICINE, 2019 (PubMed: 31645123) [IF=5.7]
8). Exosomes secreted from osteocalcin-overexpressed endothelial progenitor cells promoted endothelial cell angiogenesis. AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2019 (PubMed: 31411920) [IF=5.5]

Application: WB    Species: rat    Sample: RAOECs

Figure 2 |OCN-Exos increased endothelial cell proliferation, migration and NO formation. RAOECs were incubated with PBS, Exos and OCN-Exos for 48, and 72 hours and the proliferation abilities were detected by CCK-8 assay (A). The effects of exosomes in RAOECs migration were studied with scratch‐wound assay. After incubated with PBS, Exos and OCN-Exos for 0, 16, and 24 hours, the pictures were taken and the distance between the mean edges of the longest and shortest migration of each side were measured (C and D). eNOS and p-eNOS protein levels were analysed by western blot and the phosphorylation rate were calculated by p-eNOS/ eNOS ×100% (E and F).
9). Cyclocarya paliurus triterpenoids attenuate glomerular endothelial injury in the diabetic rats via ROCK pathway. JOURNAL OF ETHNOPHARMACOLOGY, 2022 (PubMed: 35219820) [IF=5.4]
10). Free fatty acids induce coronary microvascular dysfunction via inhibition of the AMPK/KLF2/eNOS signaling pathway. International Journal of Molecular Medicine, 2023 (PubMed: 36928181) [IF=5.4]
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