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    enos-antibody-ab5589.pdf

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Neuroscience Neurotransmission Nitric Oxide NOS
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Anti-eNOS antibody (ab5589)

  • Datasheet
  • SDS
Reviews (8)Q&A (26)References (73)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)
  • Western blot - Anti-eNOS antibody (ab5589)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)

Key features and details

  • Rabbit polyclonal to eNOS
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

You may also be interested in

Protein
Product image
Recombinant Human eNOS protein (ab112329)
Primary
Product image
Anti-eNOS (phospho S1177) antibody (ab184154)
Primary
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HRP Anti-Interferon gamma antibody [EPR1108] (ab194149)

View more associated products

Overview

  • Product name

    Anti-eNOS antibody
    See all eNOS primary antibodies
  • 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 & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human eNOS aa 1179-1194.
    Sequence:

    SLQERQLRGAVPWAFD

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Nitric Oxide
    • NOS
    • Cardiovascular
    • Blood
    • Platelets
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cardiovascular
    • Vasculature
    • Vasodilation
    • Nitric oxide associated
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human eNOS protein (ab112329)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Human
WB
Human
Application Abreviews Notes
IHC-P
1/100. Antigen retrieval is not essential but may optimise staining.
WB (5)
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.

Notes
IHC-P
1/100. Antigen retrieval is not essential but may optimise staining.
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.
  • Target information above from: UniProt accession P29474 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4846 Human
    • Entrez Gene: 18127 Mouse
    • Entrez Gene: 24600 Rat
    • Omim: 163729 Human
    • SwissProt: P29474 Human
    • SwissProt: P70313 Mouse
    • SwissProt: Q62600 Rat
    • Unigene: 647092 Human
    • Unigene: 258415 Mouse
    • Unigene: 44265 Rat
    see all
  • 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)

    Immunolocalization of eNOS in human lung tissue using ab5589.

  • Western blot - Anti-eNOS antibody (ab5589)
    Western blot - Anti-eNOS antibody (ab5589)
    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 (Human umbilical vein endothelial cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lanes 3 & 7 & 11 : Mouse placenta tissue lysate
    Lanes 4 & 8 & 12 & 16 : Rat placenta tissue lysate
    Lanes 5 & 9 & 13 : HUVEC whole cell lysate
    Lanes 6 & 10 & 14 : RAW 264.7 whole cell lysate
    Lane 15 : Mouse 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?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody (ab5589)

    The specificity of aab5589 was demonstrated using naturally occurring variable expression to confirm specificity. Immunohistochemistry analysis of human tissue sections using this antibody showed eNOS is strongly stained in human spleen tissue and no staining in human tonsil tissue which does not express eNOS. 

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (73)

    Publishing research using ab5589? Please let us know so that we can cite the reference in this datasheet.

    ab5589 has been referenced in 73 publications.

    • Miguel-Jiménez S  et al. Does Melatonin Exert Its Effect on Ram Sperm Capacitation Through Nitric Oxide Synthase Regulation? Int J Mol Sci 21:N/A (2020). PubMed: 32197481
    • Dong W  et al. Mesenchymal-endothelial transition-derived cells as a potential new regulatory target for cardiac hypertrophy. Sci Rep 10:6652 (2020). PubMed: 32313043
    • Su E  et al. Aggravated endothelial endocrine dysfunction and intimal thickening of renal artery in high-fat diet-induced obese pigs following renal denervation. BMC Cardiovasc Disord 20:176 (2020). PubMed: 32295540
    • Liu A  et al. Metformin Delays the Development of Atherosclerosis in Type 1 Diabetes Mellitus via the Methylglyoxal Pathway. Diabetes Ther 11:633-642 (2020). PubMed: 31955370
    • Jin B  et al. Yangjing Capsule Can Improve the Function of the Testicular Angiogenesis through Activating VEGFA/eNOS Signaling Pathway. Evid Based Complement Alternat Med 2020:1957267 (2020). PubMed: 32382279
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 34 Abreviews or Q&A

    Question

    We want to perform western blotting on HUVECs (Human Umbilical Endothelial Vein Cells) and A7R5 cells lines (rat aortic smooth muscle cell), both of the cell lines contain eNOS. As we found various sources of anti-eNOS antibody for WB on Abcam webpage, such as ab66127, ab5589 and ab51086, we are wondering which one could be suitable for our needs?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 28 2013

    Answer

    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] (ab50010)

    Out of these antibodies, I would suggest using ab5589 as we have had one customer who used this in western blot and obtained good results. In addition, this antibody has also been extensively used in WB in several publications.

    Read More

    Abcam Scientific Support

    Answered on Mar 28 2013

    Immunohistochemistry (Frozen sections) abreview for Anti-eNOS antibody

    Good
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Sheep Tissue sections (carotid artery)
    Permeabilization
    Yes - PBS-tween
    Specification
    carotid artery
    Blocking step
    Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 20 2019

    Western blot abreview for Anti-eNOS antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 19 2017

    Western blot abreview for Anti-eNOS antibody

    Average
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 24 2015

    Western blot abreview for Anti-eNOS antibody

    Excellent
    Abreviews
    Abreviews
    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
    Read More

    Vidhya Ramaswamy

    Verified customer

    Submitted Sep 30 2014

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-eNOS antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Sulei Xu

    Verified customer

    Submitted Nov 13 2013

    Western blot abreview for Anti-eNOS antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 12 2013

    Question

    Band around 25-30 kDa in Western blot instead of around 130-140 kDa as expected.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 07 2012

    Answer

    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 results after trying this antibody, and let me know if there is anything else that we can do for you. I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on Nov 07 2012

    Question

    Looking for antibodies to use in ICC with human cells.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 16 2012

    Answer

    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 you have any questions or need anything else, please don't hesitate to ask.

    Have a great day!

    Read More

    Abcam Scientific Support

    Answered on Jul 16 2012

    Question

    phone call: searching antibody for eNOS and phospho eNOS on rat and mouse tissue, on IHC-P and WB.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 10 2012

    Answer

    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 against total eNOS which have been tested in mouse and rat, as well as in IHC-P and Western blot. These antibodies are therefore guaranteed by us for this use.
    https://www.abcam.com/eNOS-antibody-ab66127.html (or use the following: https://www.abcam.com/eNOS-antibody-ab66127.html).
    https://www.abcam.com/eNOS-antibody-ab5589.html (or use the following: https://www.abcam.com/eNOS-antibody-ab5589.html).

    As an antibody against phsopho eNOS, we do have the ab75639 which recognizes the S1177 on eNOS. This antibody is again tested and guaranteed in mouse, rat and IHC-P and Western blot.
    https://www.abcam.com/eNOS-phospho-S1177-antibody-ab75639.html (or use the following: https://www.abcam.com/eNOS-phospho-S1177-antibody-ab75639.html).

    I have followed up with the laboratory in regards to the protocols used for the ab75639. For the Western blot, the cell lysates have been prepared with protease and phosphatase inhibitors. Before loading, the lysates have been denatured and reduced. Importantly, the membranes were blocked with 5% BSA in TBS-T. The IHC-P has been performed with heat induced antigen retrieval in Citrate buffer pH 6.

    I hope this information is still useful. Please do not hesitate to let me know if you have any question or if you require any other information from our side in regards to these or other antibodies.

    Read More

    Abcam Scientific Support

    Answered on Jul 10 2012

    1-10 of 34 Abreviews or Q&A

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