• Product nameAnti-eNOS antibody
    See all eNOS primary antibodies
  • Description
    Rabbit polyclonal to eNOS
  • SpecificityThis antibody does not detect human inducible NOS or rat brain nNOS. ab5589 has been successfully used in Western blot and immunohistochemistry (paraffin and frozen) procedures. By Western blot, this antibody detects an ~140 kDa protein representing eNOS from human endothelial cells. Immunohistochemical staining of eNOS in Human vascular endothelial cells with ab5589 results in prominent staining of the epithelial cells.
  • Tested applicationsSuitable for: ICC/IF, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Rabbit, Guinea pig, Cow, Dog, Human, Pig
  • Immunogen

    Synthetic peptide corresponding to Human eNOS aa 1179-1194.


  • Positive control
    • IHC: Human endothelial cells. WB:canine cardiac microsomes and rat or human lung lysate.
  • General notes

    We have received mixed feedback from customers regarding rat specificity; this is why we are temporarily removing Rat from tested species list and no longer guarantee it. Our laboratory is currently testing new lots so please contact our scientific support team (technical@abcam.com) for more information.



Our Abpromise guarantee covers the use of ab5589 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
ICC/IF Use at an assay dependent concentration. PubMed: 16797727
IHC-P 1/100. Antigen retrieval is not essential but may optimise staining.
IHC-Fr 1/100.
WB 1/1000. Predicted molecular weight: 133 kDa.

By Western blot ab5589 also detects eNOS in canine cardiac microsomes.
5% BSA in TBST is recommended as blocking buffer.


  • FunctionProduces 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 specificityPlatelets, placenta, liver and kidney.
  • Involvement in diseaseVariation in NOS3 seem to be associated with susceptibility to coronary spasm.
  • Sequence similaritiesBelongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • Post-translational
    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.
  • Cellular localizationCell 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
  • Database links
  • Alternative names
    • 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
    see all

Anti-eNOS antibody images

  • IHC image of eNOS staining in human placenta formalin fixed paraffin embedded tissue section*, 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 ab5589, 1/100, 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

  • Lanes 1 - 4 : Anti-eNOS antibody (ab5589) at 1/200 dilution
    Lanes 5 - 8 : Anti-eNOS antibody (ab5589) at 1/600 dilution
    Lanes 9 - 12 : Anti-eNOS antibody (ab5589) at 1/1800 dilution
    Lanes 13 - 16 : Anti-eNOS antibody (ab5589) at 1/5400 dilution

    Lane 1 : Huvec
    Lane 2 : Raw264.7
    Lane 3 : Mouse Placenta
    Lane 4 : Rat Placenta
    Lane 5 : Huvec
    Lane 6 : Raw264.7
    Lane 7 : Mouse Placenta
    Lane 8 : Rat Placenta
    Lane 9 : Huvec
    Lane 10 : Raw264.7
    Lane 11 : Mouse Placenta
    Lane 12 : Rat Placenta
    Lane 13 : Huvec
    Lane 14 : Raw264.7
    Lane 15 : Mouse Placenta
    Lane 16 : Rat Placenta

    Lysates/proteins at 10 µg per lane.

    Predicted band size : 133 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)
  • Immunolocalization of eNOS in human lung tissue using ab5589.

References for Anti-eNOS antibody (ab5589)

This product has been referenced in:
  • Shiozawa T  et al. Dimethylarginine dimethylaminohydrolase 2 promotes tumor angiogenesis in lung adenocarcinoma. Virchows Arch 468:179-90 (2016). WB, IHC-P ; Human . Read more (PubMed: 26515557) »
  • Vielma AZ  et al. Nitric Oxide Synthase 1 Modulates Basal and ß-Adrenergic-Stimulated Contractility by Rapid and Reversible Redox-Dependent S-Nitrosylation of the Heart. PLoS One 11:e0160813 (2016). Read more (PubMed: 27529477) »

See all 22 Publications for this product

Product Wall

Any of the following antibodies would be suitable as they are guaranteed to work in human, rat and in WB:

Anti-eNOS antibody (ab66127)
Anti-eNOS antibody (ab5589)
Anti-eNOS antibody (ab51086)
Anti-eNOS antibody [NOS-E1] (ab5001...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Pig Cell lysate - whole cell (endothelial cell)
Gel Running Conditions Non-reduced Denaturing (8% gel)
Loading amount 30 µg
Specification endothelial cell
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jul 24 2015

Application Western blot
Loading amount 15 µg
Gel Running Conditions Non-reduced Denaturing (7.5%)
Sample Human Cell lysate - whole cell (endothelial cells)
Specification endothelial cells
Treatment vegf stimulated
Blocking step Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Vidhya Ramaswamy

Verified customer

Submitted Sep 30 2014

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step BSA as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: 25°C
Antigen retrieval step None
Sample Rat Tissue sections (Mesenteric venule)
Specification Mesenteric venule
Permeabilization Yes - Triton X-100
Fixative Paraformaldehyde

Sulei Xu

Verified customer

Submitted Nov 13 2013

Application Western blot
Sample Mouse Cell lysate - whole cell (lung endothelial cells)
Gel Running Conditions Reduced Denaturing (gradient gel)
Loading amount 10 µg
Specification lung endothelial cells
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 12 2013

Thank you very much for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending a free of charge vial of ab66217 which should arrive tomorrow.

Please keep me updated about the resul...

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Thank you so much for your calls today and for your questions.

Please let me know if you would like to try any of the antibodies in ICC that haven't already been tested, and I will be happy to set up the testing discount program for you. If y...

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A while ago we spoke in regards to searching for the best antibodies to use for detection of eNOS and phospho eNOS on mouse and rat tissue, on paraffin sections (IHC-P)as well as in Western blot.

We have the ab66127 or the ab5589 agai...

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Thanks for your reply.

I have searched our proforma ID's and this number does not come up. I can usually find the order based on when it was placed. Do you know the date the order was placed from Sellex?

Thanks again!

Thanks for sending the email again. Not sure what is going on but the attachement still did not appear on the email. However it seems the customer is very interested in getting a replacement so I'm happy to send one.

Please let me know what a...

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