N-WASP (Ser-484/Ser-485), phospho-specific polyclonal, anti-human, mouse, rat

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. 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.

Immunogen: 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.

Specificity: 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.

Application dilution:
ELISA: 1:2000
ICC: 1:50
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 1 hour at room temperature.

Buffer/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.