Catalog # WP2201

N-WASP (Ser-484/Ser-485), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
ICC	1:50
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. In addition, these proteins bind the ARP2/3 complex, which can nucleate actin polymerization at sites that lead to branched actin structures. WASP is expressed primarily in hematopoietic cells, while its homolog N-WASP is widely expressed. These proteins have 48% identity in human with the highest homology in the functional regions of these proteins. Phosphorylation regulates the activity of both proteins. Dual phosphorylation of WASP on serine 483 and 484 by casein kinases increase the affinity for the ARP2/3 complex. Thus, dual serine phosphorylation may be important for formation of actin-based structures in various cell types.

References

Cory, G.O. et al. (2003) Mol Cell 11(5):1229-39.
Higgs, H.N. & Pollard, T.D. (2001) Annu Rev Biochem 70:649-676.
Baba, Y. et al. (1999) Blood 93:2003.

Western blot analysis of control and alkaline phosphatase-treated (AP) neonatal rat brain lysate (20 µg/lane). Blots were probed with anti-N-WASP (Lanes 1 & 2), anti-phospho-N-WASP (S484/S485) (Lanes 3 & 4), or anti-unphosphorylated-N-WASP (S484/S485) (Lanes 5 & 6).

Immunocytochemical labeling of phosphorylated and unphosphorylated N-WASP in rabbit spleen fibroblasts. The cells were probed with N-WASP (Ser-484/Ser-485) phospho-specific and N-WASP (Ser-484/Ser-485) unphosphorylated antibodies, then the antibodies were detected using appropriate secondary antibodies conjugated to Cy3. The antibodies were used in the absence (left) or presence (right) of their respective blocking peptide (WX2205 or WX2405).

Phospho-N-WASP (S484/S485) synthetic peptide (coupled to KLH) corresponding to amino acid residues around serine 484 and 485 of human N-WASP. The human WASP sequence has a similar peptide sequence surrounding serine 483 and 484.

*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 unphosphorylated-N-WASP (S484/S485) peptide then affinity purified using phospho-N-WASP (S484/S485) peptide. The antibody detects a 63 kDa* protein corresponding to the molecular mass of phosphorylated WASP on SDS-PAGE immunoblots of Jurkat cell lysate. In rat brain, A431, human endothelial and SKN-SH cells, this antibody detects a 65 kDa* protein corresponding to phosphorylated N-WASP.

*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:

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)

This kit contains: