Overview

  • Product name
    Anti-eNOS (phospho S1177) antibody
    See all eNOS primary antibodies
  • Description
    Rabbit Polyclonal to eNOS (phospho S1177)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Guinea pig, Pig
  • Immunogen

    Synthetic peptide within Human eNOS (phospho S1177). The exact sequence is proprietary.
    Database link: P29474

  • Positive control
    • THP1 whole cell lysate (untreated and treated with 100 nM PMA treatment for 6, 24 or 48 hr), Jurkat, Raji and NCI-H929 whole cell lysate; HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab184154 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
WB 1/500 - 1/3000. Predicted molecular weight: 133 kDa.
ICC/IF 1/100 - 1/1000.

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 (phospho S1177) antibody (ab184154) at 1/1000 dilution

    Lane 1 : HUVEC whole cell lysate
    Lane 2 : VEGF (ab9571) treated HUVEC whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : goat anti-rabbit antibody (green) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 133 kDa
    Additional bands at: 150 kDa (possible post-translational modification)



    2e7 HUVEC, grown in serum free medium for 18 hours, were harvested with 0.5 mM EDTA. Cells, collected and resuspended in 10 mL cell culture medium, were stimulated for 2 minutes with 50 ng per mL VEGF (ab9571), after which 30 mL of ice-cold PBS was added. The tubes were spun for 2 min at 1.5k RPM and cell pellets were snap frozen. Control cells were treated in parallel, but without the addition of VEGF.

    This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour before being incubated with anti-eNOS (phospho S1177) antibody (ab184154; diluted 1:1000) and loading control (anti-GAPDH antibody [6C5] ab8245; diluted 1:20000) at 4°C overnight. Antibody binding was detected using infrared labelled goat anti-rabbit (green) or anti-mouse (red) antibodies (diluted 1:20000) for 1 hour at room temperature before imaging.

  • All lanes : Anti-eNOS (phospho S1177) antibody (ab184154) at 1/500 dilution

    Lanes 1 & 3 & 5 : THP1 whole cell lysate
    Lane 2 : THP1 whole cell lysate (100 nM PMA treatment for 6 hr)
    Lane 4 : THP1 whole cell lysate (100 nM PMA treatment for 24 hr)
    Lane 6 : THP1 whole cell lysate (100 nM PMA treatment for 48 hr)

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 133 kDa



    5 % SDS-PAGE

  • All lanes : Anti-eNOS (phospho S1177) antibody (ab184154) at 1/500 dilution

    Lane 1 : Jurkat whole cell lysate
    Lane 2 : Raji whole cell lysate
    Lane 3 : NCI-H929 whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 133 kDa



    5 % SDS-PAGE

  • Immunofluorescent analysis of methanol fixed HeLa cells labeling eNOS using ab184154 at 1/500 dilution. Image at the right costained with Hoechst 33342.

References

This product has been referenced in:
  • Chang SH  et al. Propylthiouracil-coated biodegradable polymer inhibited neointimal formation and enhanced re-endothelialization after vascular injury. Int J Nanomedicine 13:1761-1771 (2018). Read more (PubMed: 29606869) »
  • Wang X & Song Q Mst1 regulates post-infarction cardiac injury through the JNK-Drp1-mitochondrial fission pathway. Cell Mol Biol Lett 23:21 (2018). Read more (PubMed: 29760744) »
See all 11 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (rat aorta)
Permeabilization
Yes - 0.1% triton X-100
Specification
rat aorta
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Mozhgan Shojaee

Verified customer

Submitted Aug 04 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (rat aortic endothelial cells)
Permeabilization
Yes - 0.1% triton X-100
Specification
rat aortic endothelial cells
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Mozhgan Shojaee

Verified customer

Submitted Aug 04 2017

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