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Technical Data Sheet

WK6110

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N-WASP Phospho-Regulation
Antibody Sampler Kit
Price

$395

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
WP2601 Uenishi, E. et al. (2013) J of Bio Chem 288(36): 25851. WB: MIN6-K8 -cells
WP2101 Cheney, L. et al. (2011) J Infectious Diseases. 203:1824. WB: mouse adipocytes
WP2201 Cheney, L. et al. (2011) J Infectious Diseases. 203:1824. WB: mouse adipocytes
WP2601 Cheney, L. et al. (2011) J Infectious Diseases. 203:1824. WB: mouse adipocytes
WP2001 Anitei, M. et al. (2010) Nature Cell Biol. 12:330. WB: human HeLa cells
WP2001 Hartig, S.M. et al. (2009) J Cell Sci. 122:2283. WB: rat L6 myoblasts
WP2001 Pichot, C.S. et al. (2009) British J Cancer. 1 –10. WB: breast cancer cell lines
WP2601 Pichot, C.S. et al. (2009) British J Cancer. 1 –10. WB: breast cancer cell lines
Kit Summary
The N-WASP phospho-regulation antibody sampler kit can be used to examine phosphorylation of N-WASP at Tyr-256 and Ser-484/Ser-485. For the latter site, phosphorylated relative to unphosphorylated Ser-484/Ser-485 can be monitored. In addition, the conserved sites in WASP (Tyr-291 and Ser-483/Ser-484) can also be examined. The kit includes polyclonal antibodies to monitor total expression levels of N-WASP and WASP.
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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).

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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) (WP2601) antibodies, then the antibodies were detected using appropriate secondary antibody conjugated to DyLight 594.).

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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. Dual phosphorylation of WASP on serine 483 and 484 by casein kinases increase the affinity for the ARP2/3 complex. 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.
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.
Cory, G.O. et al. (2003) Mol Cell. 11(5):1229-39.
Torres, E. & Rosen, M.K. (2003) Mol Cell 11:1215.
Wu, X. et al. (2004) J Biol Chem 279(10):9565.
Buffer and Storage
Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store all at –20°C. Stable for 1 year.
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