Phospho-NOS3 (Ser615) Antibody

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Description

Antibody Characteristics

Phospho-NOS3 (Ser615) Antibody is a rabbit-derived polyclonal antibody with high specificity for the phosphorylated form of NOS3 at Ser615. Key properties include:

ParameterDetails
Host SpeciesRabbit
ClonalityPolyclonal
ImmunogenSynthetic peptide spanning Ser615 (e.g., F-N-S(p)-I-S in humans)
Reactive SpeciesHuman, Mouse, Rat (predicted: Pig, Bovine, Horse)
ApplicationsWestern Blot (WB), ELISA, IHC, Immunofluorescence (IF)
Molecular Weight~130–133 kDa (observed via WB)
Storage-20°C; avoid freeze-thaw cycles

Biological Context

NOS3 (endothelial nitric oxide synthase) produces NO, a key vasodilator. Phosphorylation at Ser615 modulates enzymatic activity:

  • Activation Pathways: AMPK phosphorylates Ser615, enhancing NO production under calcium/calmodulin regulation .

  • Functional Impact: Phosphorylation stabilizes eNOS dimerization, critical for vascular tone and angiogenesis .

  • Disease Relevance: Dysregulation linked to ischemic stroke, hypertension, and coronary artery disease .

3.1. Recommended Dilutions

ApplicationDilution RangeSample Types
Western Blot1:500 – 1:2000Human placenta, mouse spleen, rat kidney
ELISA1:40,000Cell lysates, serum
IHC/IF1:200 – 1:1000Paraffin-embedded tissues

3.2. Validation Data

  • WB: Detects a single band at 133 kDa in human HepG2, THP-1, and mouse tissues .

  • IHC: Localizes to endothelial cells in lung cancer, spleen, and kidney tissues .

  • Flow Cytometry: Confirmed in permeabilized HepG2 cells .

Supplier Comparison

SupplierCatalog #ConcentrationImmunogen SequenceCross-Reactivity
Boster BioA01604-20.5 µg/mLQ1052-P1203 (human eNOS) Human, Mouse, Rat
Aviva Systems BioOASG051131 mg/mLS550-S630 (human) Human, Mouse, Rat
Affinity BiosciencesAF00351 mg/mLF-N-S(p)-I-S (human) Human, Mouse, Rat, Pig (predicted)

Key Research Findings

  • Vascular Studies: Used to confirm eNOS activation in VEGF-induced angiogenesis .

  • Disease Models: Detects phosphorylated eNOS in endothelial dysfunction linked to diabetes and atherosclerosis .

  • Mechanistic Insights: Phospho-Ser615 antibodies revealed AMPK’s dual role in activating/inhibiting eNOS via Thr495 phosphorylation under low calcium .

Limitations & Considerations

  • Specificity: Validated only in fixed/permeabilized cells; requires optimization for novel applications .

  • Storage: Degrades after repeated freeze-thaw cycles .

  • Research-Use Only: Not validated for diagnostic purposes .

Product Specs

Form
Rabbit IgG in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Lead Time
Typically, we can ship the products within 1-3 business days after receiving your order. Delivery times may vary depending on the purchasing method or location. Please consult your local distributors for specific delivery timeframes.
Synonyms
cNOS antibody; Constitutive NOS antibody; EC NOS antibody; EC-NOS antibody; ecNOS antibody; Endothelial nitric oxidase synthase antibody; Endothelial nitric oxide synthase antibody; Endothelial nitric oxide synthase 3 antibody; Endothelial NOS antibody; eNOS antibody; Nitric oxide synthase 3 (endothelial cell) antibody; Nitric oxide synthase 3 antibody; Nitric oxide synthase 3 endothelial cell antibody; Nitric oxide synthase endothelial antibody; Nitric oxide synthase; endothelial antibody; NOS 3 antibody; NOS III antibody; NOS type III antibody; NOS3 antibody; NOS3_HUMAN antibody; NOSIII antibody
Target Names
Uniprot No.

Target Background

Function
Endothelial nitric oxide synthase 3 (eNOS), also known as NOS3, is a critical enzyme that produces nitric oxide (NO). NO plays a vital role in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway, promoting vasodilation and contributing to blood pressure regulation. Additionally, NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels, stimulating the growth of new blood vessels. Furthermore, NO promotes blood clotting through the activation of platelets. The Phospho-NOS3 (Ser615) Antibody specifically recognizes eNOS phosphorylated at Ser615. This antibody is useful for investigating the role of Ser615 phosphorylation in the regulation of eNOS activity and NO production. This antibody also provides a tool for understanding the mechanisms involved in vascular function, angiogenesis, and platelet activation. Importantly, the antibody recognizes the inactive, dominant-negative form of eNOS. This form lacks eNOS activity and may down-regulate eNOS activity by forming heterodimers with isoform 1. This makes the antibody a valuable tool for studying the regulation of eNOS activity and the potential role of inactive eNOS in vascular disease.
Gene References Into Functions
  • The NOS3 895(G>T) polymorphism is significantly associated with recurrence-free survival in patients who received intravesical chemotherapy with pirarubicin after complete transurethral resection. PMID: 30125887
  • Human gastric cancer tissues with low BUBR1 expression showed no eNOS expression. A decrease in BUBR1 reduced eNOS bioavailability through a pathway other than eNOS phosphorylation. PMID: 30396924
  • Research suggests that overexpressing or activating eNOS in epithelial ovarian cancers (EOCs) increases their survival and enhances their capacity to regulate smooth muscle cell (SMC) migration through paracrine effects. PMID: 29343714
  • A study examining the relationship between renal colic and endothelial nitric oxide synthase gene polymorphisms found no association. PMID: 28802544
  • miR-195 and miR-582 regulate NO release by targeting the 3'-UTR of NOS3 post-transcriptionally in endothelial cells. PMID: 29948755
  • Binding of IL-5 to IL-5Ralpha receptors enhances angiogenic responses by stimulating the expression of HSP70-1 via the eNOS signaling pathway. PMID: 28317868
  • XBP1 splicing can regulate eNOS expression and cellular location, contributing to endothelial cell (EC) migration, wound healing, and angiogenesis. PMID: 29352987
  • An unfavorable genotype of the polymorphic variant of the NOS3 gene (T786C) is associated with changes in levels of active substances in individuals exposed to mercury. PMID: 30351652
  • The 4a/b polymorphism of the NOS3 gene in patients with various stages of Pneumoconiosis correlates with early development and unfavorable course of Pneumoconiosis in the post-contact period. PMID: 30351692
  • Infants with genotype GT eNOS 894G > T have a 3.4-fold higher risk of developing intraventricular hemorrhage (IVH) if they are born before 28 + 6 weeks of gestation. PMID: 28211916
  • Physical interaction between p38 and eNOS has been demonstrated, suggesting a novel, NO-independent mechanism for eNOS regulation of TLR4. Biopsy samples in patients with systemic lupus erythematosus showed reduced eNOS expression with associated elevations in TLR4 and p38, suggesting an in vivo link. PMID: 29061842
  • A negative regulatory association exists between miR24 and NOS3. Downregulation of NOS3 may induce vasospasm following subarachnoid hemorrhage, which may be due to the upregulation of miR24 in vascular smooth muscle cells (VSMCs). PMID: 29845232
  • A significant relationship was found between eNOS gene polymorphisms and the occurrence of congenital heart defects in patients with Down syndrome. Screening for the presence or absence of eNOS polymorphisms may be useful for obtaining preliminary data on the risk of congenital heart defects in patients with Down syndrome. PMID: 30204958
  • Common genetic polymorphisms in the eNOS gene contribute to the risk of erectile dysfunction, likely through effects on eNOS activity and NO availability. PMID: 29654965
  • ZYZ-803 stimulated the expression of cystathionine gamma-lyase (CSE) for H2S generation and the activity of endothelial NO synthase (eNOS) for NO production. Blocking CSE and/or eNOS suppressed ZYZ-803-induced H2S and NO production and cardioprotection. PMID: 29288927
  • Meta-analysis found that the eNOS CC genotype was not related to higher susceptibility of migraine compared with TT+ TC genotypes. Subgroup analysis showed that the CC variant increased the risk for migraine compared with TT+ TC genotypes in Caucasian populations, but this was not observed in non-Caucasian populations. No significant difference was found for other genotypes and alleles between migraine patients and healthy controls. PMID: 30200152
  • Patients with type 2 diabetes (T2D) with different genotypes at CD36, NOS3, and PPARG respond differentially to omega-3 supplements in terms of blood lipid profiles. PMID: 29703528
  • The effect of statins on the expression of sirtuin 1 (SIRT1) and endothelial nitric oxide synthase 3 (eNOS) proteins was investigated in young premature myocardial infarction (PMI) patients. Patients with PMI who were taking statins had a markedly higher level of SIRT1 compared with controls. The level of eNOS protein was considerably lower in PMI patients compared with the control group. PMID: 29664427
  • The eNOS-Glu298Asp variant (in mothers and newborns) in association with dyslipidemia (increased cholesterol, LDL, and TG levels, and decreased HDL levels) could affect NO bioavailability and represent an increased risk for preeclampsia. PMID: 28486825
  • Increased levels of nitric oxide in men with arterial hypertension did not depend on polymorphic genotypes GG and GT of the eNOS gene. PMID: 29658078
  • The C allele of the eNOS SNP 786 T/C rs2070744 was independently associated with an increased risk for cardiac instability following aneurysmal subarachnoid hemorrhage. PMID: 29079038
  • A reduction in eNOS and VEGF expression from baseline to the first clinical evaluation may indicate a response to bevacizumab. PMID: 28465540
  • The joint effect of polymorphisms of EDNRB and NOS3 on diabetic retinopathy risk was greater than the individual effect of each polymorphism in the analyzed genetic models. PMID: 28817788
  • Polymorphisms in the eNOS "A/A" (homozygous mutant) and ACE "I/D" genotypes might contribute to the increased risk of non-small cell lung cancer (NSCLC) in the South Indian population. PMID: 27328622
  • The eNOS G894T gene polymorphism was associated with the occurrence and development of coronary heart disease in young people. PMID: 29359785
  • The frequency of the T allele of the eNOS Gene in Type 2 Diabetes was less common than in controls. PMID: 28499789
  • A tandem repeat variant within intron 4 of the NOS3 gene is associated with an increased risk of infertility in men diagnosed with idiopathic oligoasthenozoospermia. PMID: 28466478
  • Upregulation of placenta-associated serum exosomal miR155 from patients with preeclampsia may suppress endothelial nitric oxide synthase (eNOS) expression in endothelial cells. PMID: 29328396
  • The eNOS gene SNP rs1808593 genotype may have an important role in predicting the occurrence of pediatric systemic lupus erythematosis and central nervous system complications in pSLE. PMID: 29465350
  • Findings suggest that NOS3 polymorphisms and physical training are important interacting variables to consider in evaluating redox status, nitric oxide availability and production, and blood pressure (BP) control. PMID: 29104725
  • The eNOS rs1799983 polymorphism and T rs1799983C rs2070744 haplotype might reduce the risk of immunoglobulin A nephropathy in Chinese populations. PMID: 28946141
  • A novel mechanism for regulation of eNOS uncoupling by low shear stress via autophagy-mediated eNOS phosphorylation, which is implicated in the geometrical nature of atherogenesis, has been reported. PMID: 29466710
  • NOS3 SNPs are associated with post-exercise hypotension in an ethnicity and exercise intensity-dependent manner. PMID: 29180482
  • Acidic pHi reduced NO synthesis and eNOS serine(1177) phosphorylation. Thus, system y(+)L activity is downregulated by an acidic pHi, a phenomenon that may result in reduced NO synthesis in human umbilical vein endothelial cells (HUVECs). PMID: 29410170
  • The meta-analysis did not detect any association between eNOS 27VNTR (4b/4a) polymorphism and diabetic microvascular complications susceptibility in Chinese populations. PMID: 29096758
  • Pitavastatin increases eNOS expression and inhibits LPS-induced miR-155 expression to prevent HUVEC cell inflammation. PMID: 28664667
  • The 27-bp VNTR polymorphism in intron 4 of the eNOS gene polymorphism may be a significant risk factor for systemic lupus erythematosus in South Indian subjects. PMID: 29524578
  • Research provides evidence to support the hypothesis that the eNOS -786 T>C polymorphism and the -786C-4a-894G haplotype are associated with a high risk of recurrent pregnancy loss. PMID: 28605668
  • The 6-Gin attenuated the injury of HUVECs induced by high glucose (HG) through the activation of the PI3K-AKT-eNOS signal pathway. PMID: 28709132
  • Two single nucleotide polymorphisms in the eNOS gene, G894T and T-786C, are strongly associated with the risk of erectile dysfunction (Meta-Analysis). PMID: 26908069
  • Extracellular histones disarrange vasoactive mediators release through a COX1-COX2-eNOS interaction in human endothelial cells. PMID: 28244682
  • The rs1799983 NOS3 polymorphism could be associated with hypertension and diastolic blood pressure among Southern Europeans. This association is influenced by dietary fat (saturated fatty acids and monounsaturated fatty acids) and body mass index. PMID: 26994605
  • The T786C eNOS mutation is common among patients with primary osteonecrosis. PMID: 28877324
  • Mechanical perturbations sensitize human red blood cell-eNOS to produce nitric oxide. PMID: 27345770
  • This research is the first to describe the effects of eNOS polymorphisms on different forms of sickle cell disease (SCD), the first to enroll SCD patients of Caucasian origin, and the first to determine eNOS mRNA levels in peripheral blood from steady-state SCD patients. PMID: 27871907
  • The development of cholangiocarcinoma (CCA) involves upregulation of eNOS and P-eNOS and their regulators. This may drive angiogenesis and metastasis in CCA. PMID: 27143607
  • No statistically significant correlation existed between serum level of PIN1 and systolic and diastolic blood pressure, between serum level of eNOS and diastolic blood pressure in normotensive Alzheimer's disease patients, between serum levels of PIN1, eNOS, and systolic blood pressure, and between serum eNOS and systolic and diastolic blood pressure in patients with hypertension. PMID: 28506742
  • The knockdown of VPO1 expression significantly increased serine1177 phosphorylation of eNOS, suggesting that structural changes and phosphorylation by VPO1 downregulate the expression of eNOS. PMID: 28264790
  • The -786 T/C polymorphism of the NOS3 gene is a susceptibility marker of Chronic Obstructive Pulmonary Disease (COPD) among Tunisians, correlating with nitric oxide levels and airflow obstruction. PMID: 28526204
  • Results showed that eNOS and XRCC4 VNTR variants might play a potential role in schizophrenia + nicotine dependence and/or nicotine dependence pathophysiology. PMID: 29050484

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Database Links

HGNC: 7876

OMIM: 163729

KEGG: hsa:4846

STRING: 9606.ENSP00000297494

UniGene: Hs.647092

Involvement In Disease
Variation Asp-298 in NOS3 may be associated with susceptibility to coronary spasm.
Protein Families
NOS family
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.

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