Overview

  • Product name
  • Description
    Rabbit polyclonal to eNOS
  • Host species
    Rabbit
  • Specificity
    This 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 applications
    Suitable 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.
    Sequence:

    SLQERQLRGAVPWAFD

  • 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.

Properties

Applications

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.

Target

  • 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
    modifications
    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 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
  • 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

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

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

This product has been referenced in:
  • Son HU  et al. The solid-state fermentation of Artemisia capillaris leaves with Ganoderma lucidum enhances the anti-inflammatory effects in a model of atopic dermatitis. Int J Mol Med 39:1233-1241 (2017). IHC-P ; Mouse . Read more (PubMed: 28393198) »
  • Jang H  et al. The herbal formula KH-204 is protective against erectile dysfunction by minimizing oxidative stress and improving lipid profiles in a rat model of erectile dysfunction induced by hypercholesterolaemia. BMC Complement Altern Med 17:129 (2017). Read more (PubMed: 28235412) »

See all 35 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Application
Western blot
Sample
Cow Cell lysate - whole cell (Aortic Endothelial Cells)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
10 µg
Specification
Aortic Endothelial Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Oct 19 2017

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
Username

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
Username

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
Username

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
Username

Abcam user community

Verified customer

Submitted Apr 12 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (stomach, kidney)
Specification
stomach, kidney
Fixative
Paraformaldehyde
Permeabilization
Yes - triton x-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Mrs. Nicole Isaacs

Verified customer

Submitted Apr 01 2009

Application
Western blot
Sample
Mouse Purified protein (brain)
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Mrs. Trisha Crandall

Verified customer

Submitted Aug 10 2005

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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