Catalog # WP2601

N-WASP (Tyr-256), 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 65 kDa

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.

References

Wu, X. et al. (2004) J Biol Chem 279(10):9565.
Torres, E. & Rosen, M.K. (2003) Mol Cell 11:1215.
Baba, Y. et al. (1999) Blood 93:2003.

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).

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.

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.

*For more information, see UniProt Accession O00401

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 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.

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)
Kalwa, H. & Michel, T. (2011) J Biol Chem 286:2320. (WB: bovine aortic endothelial cells)
Pichot, C.S. et al. (2009) British J Cancer. 1 10. (WB: breast cancer cell lines)
Elias, BC et al. (2015) J Cell Sci. 128(23):4293. (WB: mouse kidney duct cells)

This kit contains: