概述

  • 产品名称Anti-eNOS抗体[M221]
    参阅全部 eNOS 一抗
  • 描述
    小鼠单克隆抗体[M221] to eNOS
  • 特异性ab76198 is not predicted to react with other NOS family members due to low homology.
  • 经测试应用适用于: IHC-P, Flow Cyt, WB, ELISA, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Recombinant fragment of mouse eNOS protein that included amino acid residues in the C terminal region.

  • 阳性对照
    • Human umbilical vein endothelial cells untreated and treated with lambda phosphatase. Mouse placenta lysate. Huvec lysate. IHC-P: FFPE human normal placenta tissue sections.

性能

应用

Our Abpromise guarantee covers the use of ab76198 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-P 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt Use 2µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB 1/500 - 1/1000. Predicted molecular weight: 133 kDa.
ELISA 1/2000.
ICC Use at an assay dependent concentration.

靶标

  • 功能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.
  • 组织特异性Platelets, placenta, liver and kidney.
  • 疾病相关Variation in NOS3 seem to be associated with susceptibility to coronary spasm.
  • 序列相似性Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • 翻译后修饰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.
  • 细胞定位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
  • 数据库链接
  • 别名
    • 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

Anti-eNOS antibody [M221] 图像

  • All lanes : Anti-eNOS antibody [M221] (ab76198) at 1/500 dilution

    Lane 1 : Huvec cell lysates
    Lane 2 : Mouse placenta lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat anti Mouse IR680 at 1/10000 dilution

    Predicted band size : 133 kDa

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab76198 overnight at 4°C. Antibody binding was detected using Goat anti Mouse IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • IHC image of eNOS staining in human normal 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 ab76198, 1/1000 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

  • ab76198 staining eNOS in human umbilical vein endothelial cells. Cells with fixed with paraformaldehyde.

     

  • Overlay histogram showing HEK293 cells stained with ab76198 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76198, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • All lanes : Anti-eNOS antibody [M221] (ab76198) at 1/1000 dilution

    Lane 1 : human umbilical vein endothelial cells, untreated
    Lane 2 : human umbilical vein endothelial cells, treated with lambda phosphatase


    Predicted band size : 133 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)

Anti-eNOS antibody [M221] (ab76198)参考文献

This product has been referenced in:
  • You H  et al. Diabetic nephropathy is resistant to oral L-arginine or L-citrulline supplementation. Am J Physiol Renal Physiol 307:F1292-301 (2014). Read more (PubMed: 25320354) »
  • Sakellariou GK  et al. Neuron-specific expression of CuZnSOD prevents the loss of muscle mass and function that occurs in homozygous CuZnSOD-knockout mice. FASEB J 28:1666-81 (2014). WB ; Mouse . Read more (PubMed: 24378874) »

See all 6 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (8%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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提交于 Feb 09 2016

Application Western blot
Sample Rat Tissue lysate - whole (Heart)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 50 µg
Specification Heart
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Jan 26 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (skeletal muscle (plantaris))
Gel Running Conditions Reduced Denaturing (8%)
Loading amount 40 µg
Specification skeletal muscle (plantaris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Dec 04 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (8%)
Sample Rat Tissue lysate - whole (Skeletal muscle)
Specification Skeletal muscle
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jan 28 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Tissue lysate - whole (Rat brain extract)
Specification Rat brain extract
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Jun 23 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (8% Gel)
Sample Human Tissue lysate - whole (Vastus lateralis muscle)
Specification Vastus lateralis muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Feb 18 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (8% Gel)
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Specification Skeletal muscle
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

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提交于 Jan 09 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Skeletal muscle)
Loading amount 20 µg
Specification Skeletal muscle
Gel Running Conditions Reduced Denaturing (8% Gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Dr. G.K Sakellariou

Verified customer

提交于 May 15 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application Flow Cytometry
Sample Rat Cell (Aortic Endothelial Cell)
Specification Aortic Endothelial Cell
Fixation Formaldehyde
Permeabilization Yes - 0.1% Triton
Username

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提交于 Mar 18 2013


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