Anti-eNOS 抗体 (ab5589)

Reviews (8)Q&A (26)References (73)
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

製品の概要

  • 製品名

    Anti-eNOS antibody
    eNOS 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to eNOS

  • 由来種

    Rabbit

  • 特異性

    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.

  • アプリケーション

    適用あり: IHC-P, WBmore details

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

    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

  • 特記事項

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    Platelets, placenta, liver and kidney.

  • 関連疾患

    Variation in NOS3 seem to be associated with susceptibility to coronary spasm.

  • 配列類似性

    Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.

  • 翻訳後修飾

    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.

  • 細胞内局在

    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

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • 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. 

参考文献 (73)

ab5589 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab5589 は 73 報の論文で使用されています。

  • 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

レビューと Q&A

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レビューを送る 質問を送る

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?

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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.

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Immunohistochemistry (Frozen sections) abreview for Anti-eNOS antibody

 Good
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

投稿 Feb 20 2019

Western blot abreview for Anti-eNOS antibody

 Good
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

投稿 Oct 19 2017

Western blot abreview for Anti-eNOS antibody

 Average
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

投稿 Jul 24 2015

Western blot abreview for Anti-eNOS antibody

 Excellent
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

投稿 Sep 30 2014

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

 Excellent
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

投稿 Nov 13 2013

Western blot abreview for Anti-eNOS antibody

 Good
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

投稿 Apr 12 2013

Question

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

Read More
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

Question

Looking for antibodies to use in ICC with human cells.

Read More
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

Question

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

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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.

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1-10 of 34 Abreviews or Q&A

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