Anti-eNOS antibody [M221] (ab76198)
![Western blot - Anti-eNOS antibody [M221] (ab76198)](https://www.abcam.com/ps/products/76/ab76198/Images/ab76198-254561-anti-enos-antibody-m221-western-blot.jpg "Western blot - Anti-eNOS antibody [M221] (ab76198)")
Key features and details
- Mouse monoclonal [M221] to eNOS
- Suitable for: IHC-P, ICC/IF, WB, Flow Cyt (Intra)
- Reacts with: Mouse, Human
- Isotype: IgG1
Overview
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Product name
Anti-eNOS antibody [M221]
See all eNOS primary antibodies -
Description
Mouse monoclonal [M221] to eNOS -
Host species
Mouse -
Specificity
ab76198 is not predicted to react with other NOS family members due to low homology.
This antibody detects eNOS in mouse and but at a lower intensity than in human. If you are working in mouse, we would recommend using no more than 1% milk as the blocking agent for optimal signal strength.
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Tested applications
Suitable for: IHC-P, ICC/IF, WB, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment corresponding to Mouse eNOS aa 1024-1202 (C terminal).
Database link: P70313 -
Positive control
- Human umbilical vein endothelial cells untreated and treated with lambda phosphatase. Mouse placenta lysate. Huvec lysate. IHC-P: FFPE human normal placenta tissue sections.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 50% Glycerol, PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
M221 -
Isotype
IgG1 -
Research areas
Associated products
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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 ab76198 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 |
|---|---|---|
| IHC-P |
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
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| ICC/IF | (1) |
Use at an assay dependent concentration.
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| WB | (11) |
1/500 - 1/1000. Predicted molecular weight: 133 kDa.
|
| Flow Cyt (Intra) |
Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
|
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
|
ICC/IF
Use at an assay dependent concentration. |
|
WB
1/500 - 1/1000. Predicted molecular weight: 133 kDa. |
|
Flow Cyt (Intra)
Use 2µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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 antibody [M221] (ab76198)All lanes : Anti-eNOS antibody [M221] (ab76198) at 1/500 dilution
Lane 1 : Huvec cell lysates
Lane 2 : Mouse placenta lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti Mouse IR680 at 1/10000 dilution
Predicted band size: 133 kDaThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab76198 overnight at 4°C. Antibody binding was detected using Goat anti Mouse IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [M221] (ab76198)](https://www.abcam.com/ps/products/76/ab76198/Images/ab76198-251250-anti-enos-antibody-m221-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [M221] (ab76198)IHC image of eNOS staining in human normal placenta*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab76198, 1/1000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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![Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [M221] (ab76198)](https://www.abcam.com/ps/products/76/ab76198/Images/ab76198-269542-anti-enos-antibody-m221-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [M221] (ab76198)ab76198 staining eNOS in human umbilical vein endothelial cells. Cells with fixed with paraformaldehyde.
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![Flow Cytometry (Intracellular) - Anti-eNOS antibody [M221] (ab76198)](https://www.abcam.com/ps/products/76/ab76198/Images/ab76198-91556-anti-enos-antibody-m221-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-eNOS antibody [M221] (ab76198)Overlay histogram showing HEK293 cells stained with ab76198 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76198, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions. -
![Western blot - Anti-eNOS antibody [M221] (ab76198)](https://www.abcam.com/ps/products/76/ab76198/Images/ab76198-91557-anti-enos-antibody-m221-western-blot.gif)
Western blot - Anti-eNOS antibody [M221] (ab76198)All lanes : Anti-eNOS antibody [M221] (ab76198) at 1/1000 dilution
Lane 1 : human umbilical vein endothelial
cells, untreated
Lane 2 : human umbilical vein endothelial
cells, treated with lambda phosphatase
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (108)
ab76198 has been referenced in 108 publications.
- Wee CL et al. Calcitriol Supplementation Ameliorates Microvascular Endothelial Dysfunction in Vitamin D-Deficient Diabetic Rats by Upregulating the Vascular eNOS Protein Expression and Reducing Oxidative Stress. Oxid Med Cell Longev 2021:3109294 (2021). PubMed: 33623633
- Liang L et al. Exosomes derived from miR-301a-3p-overexpressing adipose-derived mesenchymal stem cells reverse hypoxia-induced erectile dysfunction in rat models. Stem Cell Res Ther 12:87 (2021). PubMed: 33494812
- Wang JS et al. Potential mechanism of Achyranthis bidentatae radix plus semen vaccariae granules in the treatment of diabetes mellitus-induced erectile dysfunction in rats utilizing combined experimental model and network pharmacology. Pharm Biol 59:547-556 (2021). PubMed: 33962551
- Gan YR et al. Dickkopf-1/cysteine-rich angiogenic inducer 61 axis mediates palmitic acid-induced inflammation and apoptosis of vascular endothelial cells. Mol Med Rep 23:N/A (2021). PubMed: 33300071
- Lee-Huang S et al. Endothelial Nitric Oxide Synthase Knockdown in Human Stem Cells Impacts Mitochondrial Biogenesis and Adipogenesis: Live-Cell Real-Time Fluorescence Imaging. J Clin Med 10:N/A (2021). PubMed: 33562279