Catalog # VP2781

VASP (Thr-278), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
ICC	1:100
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 50 kDa

Actin filament tethering and bundling are important mechanisms involved in actin superstructure assembly. The ENA/VASP family includes VASP, mena, and Ena-Vasp-like (EVL). These multidomain proteins localize to the leading edge of filopodia where they associate with AFs, interact with profilin, and compete with capping proteins at the barbed end of AFs. Artificial relocalization of VASP from the plasma membrane to mitochondrial membranes inhibits filopodial formation and axon branching, while deletion of all three ENA/VASP proteins produces defects in cortical axon-tract formation. Regulation of VASP protein activity occurs through phosphorylation at Ser-157, Ser-239, and Thr-278. AMPK phosphorylates Thr-278, leading to impaired actin stress fiber assembly and changes in cell morphology.

References

Applewhite, D.A. et al. (2007). Mol Biol Cell. 18(7):2579.
Blume, C. et al. (2007) J Biol Chem. 282(7):4601.
Krause, M. et al. (2003) Annu Rev Cell Dev Biol. 19: 541.

Western blot image of human A431 cells stimulated with calyculin A (100 nM) for 30 min. The blots were untreated (lanes 1 & 3) or treated with lambda phosphatase (lanes 2 & 4), then probed with mouse monoclonal VASP (C-term.) antibody (lanes 1 & 2) or rabbit polyclonal VASP (Thr-278) phospho-specific antibody (lanes 3 & 4).

Immunocytochemical labeling of VASP phosphorylation in rabbit spleen fibroblasts treated with Calyculin A. The cells were labeled with rabbit polyclonal VASP (Thr-278) antibody, then detected using appropriate secondary antibodies conjugated to Cy3. The antibody was used in the absence (top left) or presence (top right) of blocking peptide (VX2785). Corresponding phase images are shown bottom left and right.

Phospho-VASP (Thr-278) synthetic peptide (coupled to carrier protein) corresponding to amino acids surrounding threonine 278 in human VASP. This sequence has significant homology to the conserved site in rat and mouse VASP, but is not conserved in mena or EVL proteins.

*For more information, see UniProt Accession P50552

Rabbit polyclonal, affinity-purified antibody is supplied in 100μl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody was affinity purified using phospho-VASP (Thr-278) peptide. The purified antibody detects a 50 kDa* band corresponding to VASP in Western blots of human A431 and HeLa cells, as well as mouse C2C12 cells that have been treated with calyculin A.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

Stone, JD et al. (2016) J Non Invasive Vasc Invest. 1:2-9. (FC: rat ATr5 smooth muscle cells)
Tojkander, S. et al. (2015) Elife. Dec. 10, 4:e06126. (WB: human osteosarcoma)
Ke, Y. et al. (2015) Cell Microbiol. 17(4):473 (WB, IF: HEK293T)
Hocking, K. et al. (2013) PLoS ONE 8(4): e60986. (WB: porcine smooth muscle)
Kim, H.R. et al. (2010) AJP Cell Phys. 298:559. (WB: ferret dVSMCs)

This kit contains: