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Technical Data Sheet

WP1771

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WAVE1 (Tyr-125), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
78 kDa

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Western blot of human SYF cSrc transformed cells untreated (lanes 1 & 3) or treated (lanes 2 & 4) with pervanadate (1 mM; 30 min). The blots were probed with anti-WAVE1 (N-terminal region) (lanes 1 & 2) or anti-WAVE (Tyr-125) (lanes 3 & 4).

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Immunocytochemical labeling of phosphorylated WAVE in pervanadate-treated mouse C2C12. The cells were labeled with rabbit polyclonal WAVE1 (N-terminal region) and WAVE (Tyr-125) antibodies, then the antibodies were detected using appropriate secondary antibodies conjugated to Cy3.

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Application
Dilution


ELISA
1:2000


ICC
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
The Wiskott–Aldrich syndrome protein (WASP) family is involved in various pathways that regulate actin cytoskeletal organization. This family includes WASP, N-WASP, and three WAVE/SCAR isoforms, WAVEs 1, 2, and 3. WAVE proteins play key roles in actin-mediated cell events, such as membrane ruffling and lamellipodia formation. WAVEs contain an N-terminal WAVE homology domain, a basic domain, a Proline-rich region, and carboxy terminal verprolin, cofilin, and acidic (VCA) region. WAVEs are thought to act downstream of the Rac GTPase, connecting Rac activation to induction of Arp 2/3-mediated actin polymerization. Regulation of WAVE activity can occur through tyrosine phosphorylation. Src phosphorylation of WAVE1 at Tyr-125 enhances binding to the Arp2/3 complex, and is required for WAVE inhibition of Arp2/3-mediated stress fiber formation. By contrast, WAVE2 phosphorylation of Tyr-150 by Abl may enhance Arp2/3 complex actin nucleation and microspike formation in fibroblasts. Thus, site-specific tyrosine phosphorylation may be important for controlling specific activities of WAVE proteins.


Background References
Miki, H. et al. (1999) J Biol. Chem. 274(39):27605.
Suetsugu, S. et al. (1999) Bioch. Biophs. Res. Comm. 260:296.
Leng, Y. et al. (2005) PNAS 102(4):1098.
Ardern, H. et al. (2006) Cell Motil. Cytosk. 63:6.
Immunogen
Phospho-WAVE (Tyr-125) peptide (coupled to KLH) corresponding to amino acid residues surrounding Tyr-125 in human WAVE1. This sequence has high homology with similar regions in rat and mouse WAVE1, and has less than 50% homology to similar regions in the conserved site in WAVE2 (Tyr-124) and WAVE3 (Tyr-125).
Buffer and Storage
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.
Specificity
This antibody was cross-adsorbed to phospho-WAVE (Tyr-150) and unphosphorylated WAVE (Tyr-125) peptides before affinity purification using phospho-WAVE (Tyr-125) peptide. In western blots, the antibody detects an 80 kDa* band corresponding to phosphorylated WAVE in human SYF cSrc transformed cells and mouse macrophages treated with pervanadate, but is not observed in control cells. These bands can be specifically blocked with phospho-WAVE (Tyr-125) peptide (WX1775), but not phospho-WAVE (Tyr-150) peptide (WX1825).

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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AK6390
Arp2/3 Complex Regulation Antibody Sampler Kit

WP1731
WAVE1 (N-terminal region) Rabbit Polyclonal

WP1791
WAVE2 (Central region) Rabbit Polyclonal

WP1821
WAVE2 (Tyr-150)[conserved site], phospho-specific Rabbit Polyclonal

WX1775
phospho-WAVE (Tyr-125) Blocking Peptide

WX1825
phospho-WAVE (Tyr-150) Blocking Peptide

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