Recombinant
RabMAb

Recombinant Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)

Overview

  • Product name

    Anti-eNOS (phospho S1177) antibody [EPR20991]
    See all eNOS primary antibodies
  • Description

    Rabbit monoclonal [EPR20991] to eNOS (phospho S1177)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WB, Dot blotmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human eNOS aa 1150 to the C-terminus (phospho S1177). The exact sequence is proprietary.
    Database link: P29474

  • Positive control

    • WB: HUVEC (+/- VEGF or pervanadate) whole cell lysates; bEND.3 whole cell lysate. IP: HUVEC treated with VEGF 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 ab215717 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 1/30.
WB 1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).
Dot blot 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 [EPR20991] (ab215717) at 1/1000 dilution

    Lane 1 : bEND.3 (mouse brain endothelioma cell line) whole cell lysate
    Lane 2 : bEND.3 (mouse brain endothelioma cell line) treated with alkaline phosphatase for 1 hour, whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

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


    Exposure time: 2 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Dot blot analysis of eNOS (phospho S1177) labeled with ab215717 at 1/1000 dilution.

    Lane 1: eNOS (phospho S1177) peptide.
    Lane 2: eNOS non-phospho peptide.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Exposure time: 1 minute.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717) at 1/1000 dilution

    Lane 1 : HUVEC (human umbilical vein endothelial cell line) whole cell lysate
    Lane 2 : HUVEC (human umbilical vein endothelial cell line) treated with 100 uM pervanadate for 10 min, whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    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.

    The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).

  • All lanes : Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717) at 1/1000 dilution

    Lane 1 : Untreated HUVEC (human umbilical vein endothelial cell line) whole cell lysate
    Lane 2 : HUVEC (human umbilical vein endothelial cell line) treated with 10 ng/ml VEGF for 5 min, whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    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.

    The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).

  • eNOS (phospho S1177) was immunoprecipitated from 0.35 mg of HUVEC (human umbilical vein endothelial cell line) treated with 10 ng/ml VEGF for 5 min, whole cell lysate with ab215717 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215717 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Lane 1: HUVEC treated with VEGF whole cell lysate 10 μg (Input).
    Lane 2: ab215717 IP in HUVEC treated with VEGF whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab215717 in HUVEC treated with VEGF whole cell lysate.

    Exposure time: 30 seconds.

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

    The 60 kDa band may be a cleaved form of eNOS (PMID: 16195740).

References

ab215717 has not yet been referenced specifically in any publications.

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