Overview

  • Product name

    Anti-VASP (phospho S157) antibody [5C6]
    See all VASP primary antibodies
  • Description

    Mouse monoclonal [5C6] to VASP (phospho S157)
  • Host species

    Mouse
  • Specificity

    This antibody recognizes the the ~50 kDa VASP protein phosphorylated at Ser157. It does not cross-react with non-phosphorylated VASP or with un-related phosphorylation sites.
  • Tested applications

    Suitable for: Flow Cyt, WB, IP, ELISA, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    A synthetic phosphopeptide corresponding to amino acids surrounding the Ser157 phosphorylation site of human VASP

  • Positive control

    • Vanadate-treated A431 cells

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.1% Sodium azide
  • Concentration information loading...
  • Purity

    Size exclusion
  • Purification notes

    Mouse monoclonal antibody purified from serum-free cell culture supernatant by thiophilic adsorption and size exclusion chromatography.
  • Clonality

    Monoclonal
  • Clone number

    5C6
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab58555 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
Flow Cyt Use 0.5µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

WB Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 42 kDa.
IP Use 1-10µg for 106 cells.
ELISA Use a concentration of 0.05 µg/ml.
ICC/IF Use a concentration of 1 - 10 µg/ml. For immunocytochemistry, fixation protocol may include up to 0.5% formaldehyde.

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

  • Overlay histogram showing THP1 cells stained with ab58555 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab58555, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

References

This product has been referenced in:

  • O'Banion CP  et al. Design and Profiling of a Subcellular Targeted Optogenetic cAMP-Dependent Protein Kinase. Cell Chem Biol 25:100-109.e8 (2018). Read more (PubMed: 29104065) »
  • Li XF  et al. Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1. Exp Mol Med 50:115 (2018). Read more (PubMed: 30185781) »
See all 4 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jun 16 2019

Answer

Thank you for contacting us regarding ab58555.

I have contacted the lab on your behalf, the antibody would be fine as well in a different buffer containing another stabilizing agent e.g. BSA. However this product is provided in PEG and sucrose as was found the antibody was suffering less during lyophilization.

The antibody itself is not stickier than other antibodies. It seems possible that the buffer with the PEG and sucrose is responsible of the low yield that you are getting after the conjugation. The buffer exchange method may need to be adjusted for improved performance.

We would recommend use of one of our concentration kits for performing buffer exchange with this product. Our antibody concentration and clean up kits, Ab102776 and ab102778utilizes a simple spin column to easily and quickly buffers. These should work in any buffer containing PEG and are covered by our Abpromise guarantee. Which guarantees scientific support, replacement or refund should any Abcam product not perform as indicated on the datasheet. More information about Ab102776, ab102778 and our Abpromise may be found at the link below.



Ab102776:https://www.abcam.com/Antibody-Concentration-Kit-1-column-ab102776.html

Ab102778:https://www.abcam.com/Antibody-Concentration-Kit-3-columns-ab102778.html

https://www.abcam.com/index.html?pageconfig=abpromise

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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