Phospho-eNOS (Ser1177) Antibody | Affinity Biosciences

Product:	Phospho-eNOS (Ser1177) Antibody
Catalog:	AF3247
Description:	Rabbit polyclonal antibody to Phospho-eNOS (Ser1177)
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Rabbit, Dog
Mol.Wt.:	130~160kD; 133kD(Calculated).
Uniprot:	P29474
RRID:	AB_2834673

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

Lead Time: Same day delivery

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

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: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

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.

Conjugate:

Unconjugated.

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:

P29474(NOS3_HUMAN) >>Visit HPA database.

Gene(ID):

NOS3(4846)

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:

P29474 NOS3_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

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

Sheep

Xenopus

Zebrafish

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P29474 As Substrate

P29474

Site	PTM Type	Enzyme	Source
K5	Methylation		Uniprot
T33	Phosphorylation		Uniprot
S53	Phosphorylation		Uniprot
Y81	Phosphorylation		Uniprot
C94	S-Nitrosylation		Uniprot
C99	S-Nitrosylation		Uniprot
S102	Phosphorylation		Uniprot
S114	Phosphorylation	Q00535 (CDK5)	Uniprot
C184	S-Nitrosylation		Uniprot
C201	S-Nitrosylation		Uniprot
Y210	Phosphorylation		Uniprot
C212	S-Nitrosylation		Uniprot
S260	Phosphorylation		Uniprot
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)	Uniprot
S508	Phosphorylation		Uniprot
T512	Phosphorylation		Uniprot
S615	Phosphorylation	P17612 (PRKACA) , Q9Y243 (AKT3) , P31751 (AKT2) , P31749 (AKT1)	Uniprot
S633	Phosphorylation	Q13237 (PRKG2) , P54646 (PRKAA2) , Q13131 (PRKAA1) , P17612 (PRKACA)	Uniprot
S634	Phosphorylation		Uniprot
T636	Phosphorylation		Uniprot
S638	Phosphorylation		Uniprot
Y657	Phosphorylation	Q14289 (PTK2B)	Uniprot
C661	S-Nitrosylation		Uniprot
S738	Phosphorylation		Uniprot
K773	Ubiquitination		Uniprot
C802	S-Nitrosylation		Uniprot
K834	Ubiquitination		Uniprot
S836	Phosphorylation		Uniprot
C853	S-Nitrosylation		Uniprot
T854	Phosphorylation		Uniprot
S870	Phosphorylation		Uniprot
K904	Ubiquitination		Uniprot
C976	S-Nitrosylation		Uniprot
C991	S-Nitrosylation		Uniprot
K1035	Ubiquitination		Uniprot
C1048	S-Nitrosylation		Uniprot
C1050	S-Nitrosylation		Uniprot
K1085	Ubiquitination		Uniprot
T1094	Phosphorylation		Uniprot
C1114	S-Nitrosylation		Uniprot
T1175	Phosphorylation		Uniprot
S1177	O-Glycosylation		Uniprot
S1177	Phosphorylation	P22612 (PRKACG) , Q9Y478 (PRKAB1) , P31749 (AKT1) , Q13131 (PRKAA1) , P54646 (PRKAA2) , Q13237 (PRKG2) , P17612 (PRKACA)	Uniprot
S1179	Phosphorylation	P31749 (AKT1)	Uniprot

Research Backgrounds

P29474_NOS3_HUMAN

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.

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). 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). 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]

5). Ginsenoside Re inhibits PDGF-BB-induced VSMC proliferation via the eNOS/NO/cGMP pathway. BIOMEDICINE & PHARMACOTHERAPY, 2019 (PubMed: 31082773) [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). Buxuhuayu Decoction Accelerates Angiogenesis by Activating the PI3K-Akt-eNOS Signalling Pathway in a Streptozotocin-Induced Diabetic Ulcer Rat Model. JOURNAL OF ETHNOPHARMACOLOGY, 2021 (PubMed: 33581257) [IF=5.4]

Application: WB Species: Rat Sample:

Fig. 13. The protein expressions of p-PI3K, p-AKT1, and p-eNOS by Western blot at 7 days after administration. n = 3 for each group. Quantifications of the bands (b–d) (mean±SEM). Compared with CG, *P < 0.05 and **P < 0.01; Compared with MG, #P < 0.05, ##P < 0.01, &P < 0.05 and &&P < 0.01.

10). Cyclocarya paliurus triterpenoids attenuate glomerular endothelial injury in the diabetic rats via ROCK pathway. JOURNAL OF ETHNOPHARMACOLOGY, 2022 (PubMed: 35219820) [IF=5.4]

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Phospho-eNOS (Ser1177) Antibody - #AF3247