Overview

  • Product name

    Anti-VASP (phospho S239) antibody [EPR21734]
    See all VASP primary antibodies
  • Description

    Rabbit monoclonal [EPR21734] to VASP (phospho S239)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Dot blot, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human VASP aa 200-300 (phospho S239). The exact sequence is proprietary.
    Database link: P50552

  • Positive control

    • WB: THP-1, untreated and treated with 10µM forskolin for 30 minutes, whole cell lysate. IP: THP-1 treated with 10uM forskolin for 30 minutes 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21734
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab218619 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 1/1000. Detects a band of approximately 46 kDa (predicted molecular weight: 39 kDa).
Dot blot 1/1000.
IP 1/30.

Target

  • Function

    Ena/VASP proteins are actin-associated proteins involved in a range of processes dependent on cytoskeleton remodeling and cell polarity such as axon guidance, lamellipodial and filopodial dynamics, platelet activation and cell migration. VASP promotes actin filament elongation. It protects the barbed end of growing actin filaments against capping and increases the rate of actin polymerization in the presence of capping protein. VASP stimulates actin filament elongation by promoting the transfer of profilin-bound actin monomers onto the barbed end of growing actin filaments. Plays a role in actin-based mobility of Listeria monocytogenes in host cells. Regulates actin dynamics in platelets and plays an important role in regulating platelet aggregation.
  • Tissue specificity

    Highly expressed in platelets.
  • Sequence similarities

    Belongs to the Ena/VASP family.
    Contains 1 WH1 domain.
  • Domain

    The EVH2 domain is comprised of 3 regions. Block A is a thymosin-like domain required for G-actin binding. The KLKR motif within this block is essential for the G-actin binding and for actin polymerization. Block B is required for F-actin binding and subcellular location, and Block C for tetramerization.
    The WH1 domain mediates interaction with XIRP1.
  • Post-translational
    modifications

    Major substrate for cAMP-dependent (PKA) and cGMP-dependent protein kinase (PKG) in platelets. The preferred site for PKA is Ser-157, the preferred site for PKG, Ser-239. In ADP-activated platelets, phosphorylation by PKA or PKG on Ser-157 leads to fibrinogen receptor inhibition. Phosphorylation on Thr-278 requires prior phosphorylation on Ser-157 and Ser-239. In response to phorbol ester (PMA) stimulation, phosphorylated by PKC/PRKCA. In response to thrombin, phosphorylated by both PKC and ROCK1. Phosphorylation at Thr-278 by AMPK does not require prior phosphorylation at Ser-157 or Ser-239. Phosphorylation modulates F-actin binding, actin filament elongation and platelet activation. Carbon monoxide (CO) promotes phosphorylation at Ser-157, while nitric oxide (NO) promotes phosphorylation at Ser-157, but also at Ser-239. Response to NO and CO is blunted in platelets from diabetic patients, and VASP is not phosphorylated efficiently at Ser-157 and Ser-239.
  • Cellular localization

    Cytoplasm. Cytoplasm > cytoskeleton. Cell junction > focal adhesion. Cell projection > lamellipodium membrane. Cell projection > filopodium membrane. Targeted to stress fibers and focal adhesions through interaction with a number of proteins including MRL family members. Localizes to the plasma membrane in protruding lamellipodia and filopodial tips. Stimulation by thrombin or PMA, also translocates VASP to focal adhesions. Localized along the sides of actin filaments throughout the peripheral cytoplasm under basal conditions.
  • Information by UniProt
  • Database links

  • Alternative names

    • Vasodilator stimulated phosphoprotein antibody
    • Vasodilator-stimulated phosphoprotein antibody
    • VASP antibody
    • VASP_HUMAN antibody

Images

  • All lanes : Anti-VASP (phospho S239) antibody [EPR21734] (ab218619) at 1/1000 dilution

    Lane 1 : Untreated THP-1 (human monocytic leukemia cell line) whole cell lysate
    Lane 2 : THP-1 treated with 10µM forskolin for 30 minutes, 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

    Predicted band size: 39 kDa
    Observed band size: 46 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST 

    This blot was developed using a higher sensitivity ECL substrate.

  • VASP (phospho S239) was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia cell line) treated with 10µM forskolin for 30min whole cell lysate using ab218619 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab218619 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.

    Lane 1: THP-1 treated with 10µM forskolin for 30min whole cell lysate 10μg (input).
    Lane 2: ab218619 IP in THP-1 treated with 10µM forskolin for 30min whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab218619 in THP-1 treated with 10µM forskolin for 30min whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

  • Dot blot analysis of VASP (phospho S239) using ab218619 at 1/1000 dilution, followed byGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.

    Lane 1: VASP (phospho S239) peptide
    Lane 2: VASP non-phospho peptide.

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

    Exposure time: 58 seconds.

References

ab218619 has not yet been referenced specifically in any publications.

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