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重组RabMAb

重组Anti-eNOS抗体[EPR23750-3] - BSA and Azide free (ab273574)

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Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
  • Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23750-3] to eNOS - BSA and Azide free
  • Suitable for: ICC/IF, IHC-P, IP, Flow Cyt (Intra), mIHC, WB
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free Carrier Free Unconjugated Unconjugated Unconjugated

概述

  • 产品名称

    Anti-eNOS抗体[EPR23750-3] - BSA and Azide free
    参阅全部 eNOS 一抗

  • 描述

    兔单克隆抗体[EPR23750-3] to eNOS - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC/IF, IHC-P, IP, Flow Cyt (Intra), mIHC, WBmore details

  • 种属反应性

    与反应: Human

  • 免疫原

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

  • 阳性对照

    • WB: EA.hy926 and HUVEC whole cell lysates. IHC-P: Human lung carcinoma, kidney, spleen and placenta tissue. ICC/IF: HUVEC cells. Flow Cyt (intra): HeLa cells. IP: EA.hy926 whole cell lysate. mIHC: Human liver tissue.

  • 常规说明

    ab273574 is the carrier-free version of ab252439.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab273574于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
mIHC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).
说明
ICC/IF
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
mIHC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).

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

    图片

    • Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      Fluorescence multiplex immunohistochemical analysis of human liver (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-eNOS stained on endothelial cells (ab252439; red; Opal™570) at 1:1000 ( 1.004 μg/ml) [Panel B], anti-CD163 stained on Kupffer cells (ab213612; green; Opal™520) at 1:8000 ( 0.13 μg/ml) [Panel B], and anti-Glucose Transporter GLUT2 stained on membrane of hepatocytes (ab234440; gray; Opal™690) at 1:200 ( 3.005 μg/ml) [Panel D] on human liver. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab234440, ab213612, and ab252439 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.

      This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    • Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      Fluorescence multiplex immunohistochemical analysis of human liver (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-MASP2 stained on cytoplasm of hepatocytes (ab277520; gray; Opal™690) at 1:100 ( 5.22 μg/ml) [Panel B] , anti-CD163 stained on Kupffer cells (ab213612; green; Opal™520) at 1:8000 ( 0.13 μg/ml) [Panel C], and anti-eNOS tained on endothelial cells (ab252439; red; Opal™570) at 1:200 ( 3.005 μg/ml) [Panel D] on human liver. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab277520, ab213612, and ab252439 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.

      This data was developed using ab252439, the same antibody clone in a different buffer formulation.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      This data was developed using ab252439, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on endothelial cells in human spleen. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      This data was developed using ab252439, the same antibody clone in a different buffer formulation.

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC cells labelling eNOS with ab252439 at 1/100 (5.99 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HUVEC cells. Negative control: HeLa (PMID: 19559671). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

    • Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      This data was developed using ab252439, the same antibody clone in a different buffer formulation. 

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell, left) /HUVEC (human umbilical vein endothelial cell, right) cells labelling eNOS with ab252439 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). 

      A Goat anti rabbit IgG (Alexa Fluor®488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

      Negative control:Hela (PMID: 19559671). 

       

       

    • Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      This data was developed using ab252439, the same antibody clone in a different buffer formulation.

      eNOS was immunoprecipitated from 0.35 mg EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate with ab252439 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252439 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate 10 ug

      Lane 2: ab252439 IP in EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252439 in EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 6 seconds.

      IP lysates were unboiled.

    • Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Anti-eNOS antibody [EPR23750-3] (ab252439) at 1/1000 dilution + HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

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



      This data was developed using ab252439, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      HUVEC whole cell lysate in lane 1 is made freshly and used in WB test immediately to minimize protein degradation.

      Exposure time: 6 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      This data was developed using ab252439, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling eNOS with abab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on endothelial cells in human lung carcinoma. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Western blot - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      This data was developed using ab252439, the same antibody clone in a different buffer formulation. 

      Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling eNOS with abab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on endothelial cells in human lung carcinoma. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      This data was developed using ab252439, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human placenta. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

      This data was developed using ab252439, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on endothelial cells in human kidney. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)
      Anti-eNOS antibody [EPR23750-3] - BSA and Azide free (ab273574)

    实验方案

    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

    文献 (0)

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