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Technical Data Sheet

WP2401

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unphosphorylated N-WASP (Ser-484/Ser-485)
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
65 kDa

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Enlarge

Western blot 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).

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Enlarge

Immunocytochemical labeling of phospho- and unphospho-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 blocking peptide (WX2205 or WX2405).

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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 1hour at room temperature.
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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. 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.


Background References
Baba, Y. et al. (1999) Blood 93:2003.
Higgs, H.N. & Pollard, T.D. (2001) Annu Rev Biochem 70:649-676.
Cory, G.O. et al. (2003) Mol Cell. 11(5):1229-39.
Immunogen
Unphosphorylated 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.
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-N-WASP (S484/S485) peptide (without carrier) then affinity purified using unphosphorylated N-WASP (S484/S485) peptide (without carrier). The antibody detects a 65 kDa* protein on SDS-PAGE immunoblots of rat brain and SKN-SH cell lysates that are treated with alkaline phosphatase. Only low levels of unphosphorylated WASP or N-WASP are detected in untreated rat brain, Jurkat, and A431 cells.

*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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