Overview

  • Product name
    Anti-eNOS antibody [EPR19296]
    See all eNOS primary antibodies
  • Description
    Rabbit monoclonal [EPR19296] to eNOS
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human eNOS aa 250-350. The exact sequence is proprietary.
    Database link: P29474

  • Positive control
    • WB: Human fetal heart, fetal kidney, fetal liver and placenta lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart and spleen lysates; HepG2 whole cell lysate; whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 2 hours and 6 hours; DDDDK tagged human eNOS full length recombinant protein (aa2-1203) ab198066. IP: HepG2 whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab199956 in the following tested applications.

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

Application Abreviews Notes
IP 1/50.
WB 1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).

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.
  • Information by UniProt
  • Database links
  • 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

  • All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : Human fetal liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

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


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    eNOS is shown to be expressed in platelets, placenta, liver and kidney (UniProt entry P29474). It also plays an important role in the cardiovascular system. (PMID: 15210449 & 15936740).

  • All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat heart lysate
    Lane 7 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (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?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1, 2, 3, 4, 6 and 7: 30 seconds; Lane 5: 2 minutes.

    eNOS is shown to be expressed in platelets, placenta, liver and kidney (UniProt entry P29474). It also plays an important role in the cardiovascular system. (PMID: 15210449 & 15936740).

  • All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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



    Exposure Time: Lane1: 40 seconds; Lane2: 180 seconds

    The expression profile observed in HeLa is consistent with the literature (PMID: 19925457).
    Negative control: HeLa (PMID: 19925457)

  • All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : Human placenta lysate
    Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 15 seconds; Lane 2: 1 minute.

    The expression profile observed is consistent with what has been described in the literature (PMID:17855661).

  • All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/5000 dilution

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : Whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 2 hours
    Lane 3 : Whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 6 hours

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

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


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID:17855661 & 11004215).

  • All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : Recombinant Human eNOS protein (Tagged) (ab198066) at 0.01 µg
    Lane 2 : Recombinant Human iNOS protein (ab135010) at 0.05 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (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?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 1 second; Lane 2: 3 minutes.

     

  • eNOS was immunoprecipitated from 1 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab199956 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab199956 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HepG2 whole cell lysate 10 μg (Input).

    Lane 2: ab199956 IP in HepG2 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199956 in HepG2 whole cell lysate.

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

    Exposure time: 3 seconds.

References

This product has been referenced in:
  • Wang Y & Jin L miRNA-145 is associated with spontaneous hypertension by targeting SLC7A1. Exp Ther Med 15:548-552 (2018). Read more (PubMed: 29434681) »
  • Tu F  et al. Angiogenic effects of apigenin on endothelial cells after hypoxia-reoxygenation via the caveolin-1 pathway. Int J Mol Med 40:1639-1648 (2017). Read more (PubMed: 29039442) »
See all 2 Publications for this product

Customer reviews and Q&As

Application
Immunoprecipitation
Sample
Mouse Tissue lysate - whole (liver)
Total protein in input
2000 µg
Immuno-precipitation step
Other - Protein G dynabeads
Specification
liver

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Verified customer

Submitted Oct 10 2018

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