Recombinant Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)
![Western blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)](https://www.abcam.com/ps/products/215/ab215717/Images/ab215717-297339-anti-enos-phospho-s1177-antibody-epr20991-western-blot.jpg "Western blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20991] to eNOS (phospho S1177)
- Suitable for: IP, WB, Dot blot
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-eNOS (phospho S1177) antibody [EPR20991]
See all eNOS primary antibodies -
Description
Rabbit monoclonal [EPR20991] to eNOS (phospho S1177) -
Host species
Rabbit -
Tested applications
Suitable for: IP, WB, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HUVEC (+/- VEGF or pervanadate) whole cell lysates; bEND.3 whole cell lysate. IP: HUVEC treated with VEGF whole cell lysate. Dot blot: eNOS (phospho S1177) peptide.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20991 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab215717 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IP |
1/30.
|
|
| WB |
1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).
|
|
| Dot blot |
1/1000.
|
| Notes |
|---|
|
IP
1/30. |
|
WB
1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa). |
|
Dot blot
1/1000. |
Target
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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.
Isoform eNOS13C: Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1. -
Tissue specificity
Platelets, placenta, liver and kidney. -
Involvement in disease
Variation in NOS3 seem to be associated with susceptibility to coronary spasm. -
Sequence similarities
Belongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain. -
Post-translational
modificationsPhosphorylation 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. -
Cellular localization
Cell membrane. Membrane, caveola. Cytoplasm, cytoskeleton. Golgi apparatus. Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle and which is favored by interaction with NOSIP and results in a reduced enzymatic activity. - Information by UniProt
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Database links
- Entrez Gene: 4846 Human
- Entrez Gene: 18127 Mouse
- Omim: 163729 Human
- SwissProt: P29474 Human
- SwissProt: P70313 Mouse
- Unigene: 647092 Human
- Unigene: 258415 Mouse
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Alternative names
- cNOS antibody
- Constitutive NOS antibody
- EC NOS antibody
see all
Images
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Western blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)All lanes : Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717) at 1/1000 dilution
Lane 1 : bEND.3 (mouse brain endothelioma cell line) whole cell lysate
Lane 2 : bEND.3 (mouse brain endothelioma cell line) treated with alkaline phosphatase for 1 hour, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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![Dot Blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)](https://www.abcam.com/ps/products/215/ab215717/Images/ab215717-297390-anti-enos-phospho-s1177-antibody-epr20991-dot-blot.jpg)
Dot Blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)Dot blot analysis of eNOS (phospho S1177) labeled with ab215717 at 1/1000 dilution.
Lane 1: eNOS (phospho S1177) peptide.
Lane 2: eNOS non-phospho peptide.Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.
Exposure time: 1 minute.
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Immunoprecipitation - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)](https://www.abcam.com/ps/products/215/ab215717/Images/ab215717-297336-anti-enos-phospho-s1177-antibody-epr20991-immunoprecipitation.jpg)
Immunoprecipitation - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)eNOS (phospho S1177) was immunoprecipitated from 0.35 mg of HUVEC (human umbilical vein endothelial cell line) treated with 10 ng/ml VEGF for 5 min, whole cell lysate with ab215717 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215717 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: HUVEC treated with VEGF whole cell lysate 10 μg (Input).
Lane 2: ab215717 IP in HUVEC treated with VEGF whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab215717 in HUVEC treated with VEGF whole cell lysate.Exposure time: 30 seconds.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).
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![Western blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)](https://www.abcam.com/ps/products/215/ab215717/Images/ab215717-297338-anti-enos-phospho-s1177-antibody-epr20991-western-blot.jpg)
Western blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)All lanes : Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717) at 1/1000 dilution
Lane 1 : HUVEC (human umbilical vein endothelial cell line) whole cell lysate
Lane 2 : HUVEC (human umbilical vein endothelial cell line) treated with 100 uM pervanadate for 10 min, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).
-
![Western blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)](https://www.abcam.com/ps/products/215/ab215717/Images/ab215717-297337-anti-enos-phospho-s1177-antibody-epr20991-western-blot.jpg)
Western blot - Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)All lanes : Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717) at 1/1000 dilution
Lane 1 : Untreated HUVEC (human umbilical vein endothelial cell line) whole cell lysate
Lane 2 : HUVEC (human umbilical vein endothelial cell line) treated with 10 ng/ml VEGF for 5 min, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).
-
![Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)](https://www.abcam.com/ps/products/215/ab215717/Images/ab215717-6-benefits-of-recombinant-antibodies.png)
Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (19)
ab215717 has been referenced in 19 publications.
- Cai C et al. Empagliflozin attenuates cardiac microvascular ischemia/reperfusion through activating the AMPKα1/ULK1/FUNDC1/mitophagy pathway. Redox Biol 52:102288 (2022). PubMed: 35325804
- Velagic A et al. Cardioprotective actions of nitroxyl donor Angeli's salt are preserved in the diabetic heart and vasculature in the face of nitric oxide resistance. Br J Pharmacol 179:4117-4135 (2022). PubMed: 35365882
- Zhang G et al. Radix Astragalus Polysaccharide Accelerates Angiogenesis by Activating AKT/eNOS to Promote Nerve Regeneration and Functional Recovery. Front Pharmacol 13:838647 (2022). PubMed: 35431954
- Gao F et al. LncRNA HOXA11-AS promotes vascular endothelial cell injury in atherosclerosis by regulating the miR-515-5p/ROCK1 axis. ESC Heart Fail 9:2259-2271 (2022). PubMed: 35578440
- Whitehead AK et al. Alpha7 nicotinic acetylcholine receptor mediates chronic nicotine inhalation-induced cardiopulmonary dysfunction. Clin Sci (Lond) 136:973-987 (2022). PubMed: 35678315