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Technical Data Sheet

WP2601

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N-WASP (Tyr-256), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
65 kDa

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Western blot analysis of control and pervanadate-treated A431 cells (20 µg/lane). Blots were probed with either rabbit polyclonal anti-N-WASP (Ser-484/Ser-485) or anti-N-WASP (Tyr-256).

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Immunocytochemical labeling of N-WASP in control and pervanadate-treated A431 cells. The cells were labeled with rabbit polyclonal N-WASP/WASP (WP2101) or rabbit polyclonal N-WASP (Tyr-256) antibodies, then the antibodies were detected using appropriate secondary antibody conjugated to DyLight® 594.

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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.
Product References
Uenishi, E. et al. (2013) J of Bio Chem 288(36): 25851. WB: MIN6-K8 -cells
Cheney, L. et al. (2011) J Infectious Diseases. 203:1824. WB: mouse adipocytes
Pichot, C.S. et al. (2009) British J Cancer. 1 –10. WB: breast cancer cell lines
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Background
Members of the Wiskott-Aldrich sydrome protein (WASP) family regulate the formation of actin-based cell structures in many cell types. These proteins contain C-terminal actin-binding domains that can stimulate actin polymerization. WASP is expressed primarily in hematopoietic cells, while its homolog N-WASP is widely expressed. These proteins have 48% identity in human with higher homology in the functional regions of these proteins. Phosphorylation at serine and tyrosine residues regulates the activity of both proteins. WASP is tyrosine phosphorylated at tyrosine 291 after antigen receptor activation in B-cells and collagen stimulation of platelets. Phosphorylation of the analogous site in N-WASP (Tyr-256) stimulates its activity, reduces nuclear N-WASP, and is required for neurite extension.


Background References
Baba, Y. et al. (1999) Blood 93:2003.
Torres, E. & Rosen, M.K. (2003) Mol Cell 11:1215.
Wu, X. et al. (2004) J Biol Chem 279(10):9565.
Immunogen
Phospho-N-WASP (Tyr-256) synthetic peptide (coupled to BSA) corresponding to amino acid residues around tyrosine 256 of human N-WASP. The human WASP sequence has a two amino acid difference in the same region surrounding tyrosine 291.
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 phosphotyrosine then affinity purified using phospho-N-WASP (Tyr-256) peptide (without carrier). The antibody detects a 65 kDa* protein corresponding to the molecular mass of phosphorylated N-WASP on SDS-PAGE immunoblots of A431 cells treated with pervanadate. A similar band is observed in pervanadate treated HeLa and endothelial cells. Weak bands are also observed at higher molecular weights after pervanadate treatment. These bands may be due to low cross-reactivity with phosphotyrosine.

*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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