• Product nameAnti-eNOS antibody
    See all eNOS primary antibodies
  • Description
    Rabbit polyclonal to eNOS
  • SpecificityDoes not detect human inducible NOS or rat brain NOS. eNOS is expressed in only a few human tissues: platelets, placenta, liver and kidney (UniProt entry P29474). Expression levels can vary greatly. According to the Human Protein Atlas, eNOS is expressed at low levels in colon, kidney, prostate cancer, at medium levels in colorectal cancer, lung cancer and melanoma, and at high levels in placenta. Mouse eNOS: UniProt entry P70313. Rat eNOS: Expressed constitutively by vascular endothelium. Detected in alveolar and serosal epithelial cells as well as in endothelial cells in one day old rat. In adult lung, detected in rare endothelial cells. Detected at high levels in lung during the late fetal and postnatal period and at lower levels in adult (UniProt entry Q62600).
  • Tested applicationsSuitable for: WB, IP, ELISA, IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow, Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to an internal sequence of human eNOS.

  • Positive control
    • Lung. Placenta.



Our Abpromise guarantee covers the use of ab66127 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
WB 1/200. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).

In some samples containing lower levels of endogenous eNOS, IP needs to be performed to enrich samples.

Because eNOS is of higher MW (140 kDa), it is possible that the Immunoprecipitated protein does not transfer sufficiently from gel to membrane. We suggest to transfer the protein overnight at 4oC.

Blocking is recommended with 3% BSA

IP Use a concentration of 2 - 5 µg/ml.
ELISA Use a concentration of 0.01 - 0.1 µg/ml.
IHC-P 1/50 - 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. The recommended dilutions were determined for a 10-minute incubation at room temperature. Dilute further if results show high background.
ICC/IF Use a concentration of 5 µg/ml.
IHC-Fr Use at an assay dependent concentration.


  • 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*, 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 ab66127, 1/100 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

  • Anti-eNOS antibody (ab66127) at 1/450 dilution + Huvec (positive control) at 20 µg

    800CW Goat Anti-Rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 133 kDa
    Additional bands at : 140 kDa,16 kDa,32 kDa. We are unsure as to the identity of these extra bands.
  • Human placenta tissue stained with 1/50 ab66127 for 10 minutes, after antigen retrieval.
  • ICC/IF image of ab66127 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66127, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-eNOS antibody (ab66127)

This product has been referenced in:
  • Nagane M  et al. Activation of eNOS in endothelial cells exposed to ionizing radiation involves components of the DNA damage response pathway. Biochem Biophys Res Commun 456:541-6 (2015). Read more (PubMed: 25498542) »
  • Pires PW  et al. Unitary TRPV3 channel Ca2+ influx events elicit endothelium-dependent dilation of cerebral parenchymal arterioles. Am J Physiol Heart Circ Physiol 309:H2031-41 (2015). Read more (PubMed: 26453324) »

See all 14 Publications for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - whole (Valvular intestitial cells)
Gel Running Conditions Reduced Denaturing (4-12% Bis-tris)
Loading amount 40 µg
Specification Valvular intestitial cells
Blocking step Blocking buffer for WB (Rockland) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: RT°C

Dr. Stephane Zaffran

Verified customer

Submitted May 23 2016

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (liver)
Specification liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Nov 10 2014

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...

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The antibody can be diluted in 3% BSA in TBST and blocked with the same solution.

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