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mAb IE273 anti-VASP (vasodilator stimulated phosphoprotein)
- download Reprint (PDF) version of the data sheet
Background information
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VASP (vasodilator stimulated phosphoprotein) is a proline-rich protein substrate of cAMP- and cGMP-dependent protein kinases. Phosphorylation of VASP at Ser-157 causes a mobility shift in SDS gel electrophoresis from 46 to 50 kDa, which has been used as a convenient marker to monitor cyclic nucleotide-dependent protein kinase activity. VASP is the founding member of the Ena-VASP protein family, comprising the Drosophila protein Enabled (Ena), its mouse homologue Mena (mammalian Enabled), and mouse EVL (Ena-VASP-like protein). With these proteins VASP shares a conserved overall domain organization:
a) the conserved N-terminal Ena-VASP homology domain 1 (EVH1), which mediates binding to a novel proline-rich motif,
b) a more divergent proline-rich central domain (which is responsible for profilin binding), and
c) a conserved C-terminal EVH2 domain.Particularly high VASP levels are present in platelets, although VASP is expressed in a wide variety of cell types and tissues. In cultured cells, VASP is associated with focal adhesions, cell-cell contacts, microfilaments, and highly dynamic membrane regions. From in vitro binding data VASP has been suggested to link profilin to zyxin, vinculin, and the Listeria spp. surface protein ActA, respectively. Functional evidence indicates that VASP is a crucial factor involved in the enhancement of actin filament formation and the actin-dependent motility of intracellular bacterial pathogens.
- Antibody preparation and storage
- Clone: IE273, isotype IgG1 . 50 µg of purified antibody in PBS, 0.01% (w/v) NaN3. Antibody concentration: 500 µg/ml (OD280=0.7). Vials have been overfilled by 10% to ensure complete recovery of the specified amount. Short term storage at 4°C, stable for one year from date of shipment when stored at -20°C. Avoid repeated frezzing and thawing! Do not store in "frost-free" freezers.
- Antigen
- VASP purified from human platelets
- Species cross-reactivity
- human, porcine, and bovine VASP
- Specificity
- The antibody recognizes both the 46 kDa (Ser-157 dephospho) and 50 kDa (Ser-157 phospho) form of VASP.
- Applications
- Western (immuno) blotting (0.1-0.25 µg/ml; dilution 1:2000 - 1:5000), immunofluorescence of formaldehyde fixed cells (1-5 µg/ml; dilution 1:100 - 1:500), immunoprecipitation with anti-mouse secondary antibody.
All dilutions refer to the analysis of cells and tissues with intermediate to high levels of VASP expression and must be viewed as approximate. The serum should be titrated for each individual application.
- Positive control
- Human platelet protein (500 µg), supplied at 5 mg/ml in SDS-stop solution (100 mM NaCl, 73 mM Tris/HCl pH 6.7, 10 mM DTT, 8 mM EDTA, 5 % [v/v] glycerol, 2% [w/v] SDS, 10 µg/ml Bromophenol Blue sodium salt). Use 5 µl (25 µg) per lane.
- Related products
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- rabbit antiserum M4 to human VASP, 100 µl (catalog # 0010-10)
- pre-immune serum to M4, 25 µl (catalog # 0013-02)
- positive control: human platelet protein in SDS-stop solution, 500 µg (catalog # 8010-50) - References
Literature referencing anti-VASP mAb IE273General Literature on VASPPricing & Ordering
