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リコンビナントRabMAb

Anti-eNOS 抗体 [EPR26335-79] (ab300071)

Submit a review Submit a question References (8)
Western blot - Anti-eNOS antibody [EPR26335-79] (AB300071)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (AB300071)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (AB300071)
  • Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR26335-79] (AB300071)
  • Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR26335-79] (AB300071)
  • Anti-eNOS antibody [EPR26335-79] (AB300071)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR26335-79] to eNOS
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free Carrier Free Unconjugated

製品の概要

  • 製品名

    Anti-eNOS antibody [EPR26335-79]
    eNOS 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR26335-79] to eNOS

  • 由来種

    Rabbit

  • 特異性

    Unsuitable for human, we recommend ab252439 and ab199956 for human. IHC-P unsuitable for mouse.

  • アプリケーション

    適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
    適用なし: IP

  • 種交差性

    交差種: Mouse, Rat
    非交差種: Human

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: Whole cell fresh lysates: bEnd.3 (mouse brain endothelioma). IHC-P: Rat spleen, colon, and stomach tissue. ICC/IF: bEnd.3 (mouse brain endothelioma endothelialpolyoma middle T antigen transformed). Flow cyt. intr.: bEnd.3 (mouse brain endothelioma endothelialpolyoma middle T antigen transformed).

  • 特記事項

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab300071の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 132 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

This antibody is not suitable for human species in IHC-P application.
ICC/IF
1/50.
特記事項
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 132 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

This antibody is not suitable for human species in IHC-P application.
ICC/IF
1/50.
追加情報
Is unsuitable for IP.

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

    • 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

    • Western blot - Anti-eNOS antibody [EPR26335-79] (AB300071)
      Western blot - Anti-eNOS antibody [EPR26335-79] (AB300071)
      All lanes : Anti-eNOS antibody [EPR26335-79] (ab300071) at 1/1000 dilution

      Lane 1 : bEnd.3 (mouse brain endothelioma) whole cell fresh lysate
      Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell fresh lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 132 kDa
      Observed band size: 140 kDa why is the actual band size different from the predicted?



      5% NFDM/TBST was used as blocking and diluting buffer. 

      Exposure time: 

      Lane 1: 10 seconds
      Lane 2: 3 minutes

      Negative control:RAW264.7 (PMID: 18607531)
      Lysates were freshly made and used in WB test immediately to minimize protein degradation.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (AB300071)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (AB300071)

      Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling eNOS with ab300071 at 1/50 dilution (10.96 µg/ml), followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on vascular endothelial cells of rat spleen was observed. The section was incubated with ab300071 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) was used.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (AB300071)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (AB300071)

      Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling eNOS with ab300071 at 1/50 dilution (10.96 µg/ml), followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on vascular endothelial cells of rat stomach was observed. The section was incubated with ab300071 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

    • Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR26335-79] (AB300071)
      Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR26335-79] (AB300071)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized bEnd.3 (mouse brain endothelioma endothelialpolyoma middle T antigen transformed) cells lebeling eNOS with ab300071 at 1/50 dilution (10.96 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing cytoplasm staining in bEnd.3 cell line. Negative control: RAW 264.7 (PMID: 18607531).

      ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.

    • Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR26335-79] (AB300071)
      Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR26335-79] (AB300071)

      Flow cyt (Intra) analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage, Left) / bEnd.3 (Mouse brain endothelioma, Right) cells labeling eNOS with ab300071 at 1/500 dilution (0.1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: RAW 264.7 (PMID: 18607531). 

    • Anti-eNOS antibody [EPR26335-79] (AB300071)
      Anti-eNOS antibody [EPR26335-79] (AB300071)

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (8)

    ab300071 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。

    ab300071 は 8 報の論文で使用されています。

    • Yu B  et al. Extracellular vesicles engineering by silicates-activated endothelial progenitor cells for myocardial infarction treatment in male mice. Nat Commun 14:2094 (2023). PubMed: 37055411
    • Wang L  et al. Shear stress leads to the dysfunction of endothelial cells through the Cav-1-mediated KLF2/eNOS/ERK signaling pathway under physiological conditions. Open Life Sci 18:20220587 (2023). PubMed: 37077343
    • Liu B  et al. A novel mouse model of heart failure with preserved ejection fraction after chronic kidney disease induced by retinol through JAK/STAT pathway. Int J Biol Sci 19:3661-3677 (2023). PubMed: 37564202
    • Fathi FEZM  et al. Vitamin D regulates insulin and ameliorates apoptosis and oxidative stress in pancreatic tissues of rats with streptozotocin-induced diabetes. Environ Sci Pollut Res Int 29:90219-90229 (2022). PubMed: 35864405
    • Luo M  et al. Recurrent Hypoglycemia Impaired Vascular Function in Advanced T2DM Rats by Inducing Pyroptosis. Oxid Med Cell Longev 2022:7812407 (2022). PubMed: 35915611
    • Han X Inhibiting P2Y12 receptor relieves LPS-induced inflammation and endothelial dysfunction. Immun Inflamm Dis 10:e697 (2022). PubMed: 36169256
    • Wang Y  et al. Role of GADD45A in myocardial ischemia/reperfusion through mediation of the JNK/p38 MAPK and STAT3/VEGF pathways. Int J Mol Med 50:N/A (2022). PubMed: 36331027
    • Paez-Mayorga J  et al. Implantable niche with local immunosuppression for islet allotransplantation achieves type 1 diabetes reversal in rats. Nat Commun 13:7951 (2022). PubMed: 36572684

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