Synthetic peptide corresponding to aa175-190, EAPSFLSLRRGTALSN, from mouse Nck beta, conjugated to KLH. Species Sequence Homology: human
Nck is a highly conserved, oncogenic protein encoding one SH2 and three SH3 domains, the Src homology motifs found in nonreceptor tyrosine kinases, Ras GTPase-activating protein, phosphatidylinositol 3-kinase, and phospholipase C-gamma. Nck is a common target for the action of different surface receptors. Nck is widely expressed in various tissues and in cell lines from human, murine and rat origins. Nck is phosphorylated on tyrosine, serine and threonine residues in response to stimulation of EGF and PDGF in A431 and NIH 3T3 cells, respectively. Like other SH2-containing proteins, Nck is associated with tyrosine- autophosphorylated EGF or PDGF receptors via its SH2 domain. Overexpression of Nck leads to transformation of NIH 3T3 cells. The recombinant Nck precipitates endogenous WIP, a novel proline-rich protein that interacts with the Wiskott-Aldrich syndrome protein (WASP), from BJAB cell lysates. Nck binds through its second SH3 domain to WIP, and Nck binds to WIP at a site (aa321-415) that differs from the WASP-binding site (aa416-488). WIP has been shown to associate with the actin polymerization regulatory protein profilin and to induce actin polymerization and cytoskeletal reorganization in lymphoid cells. Applications: Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested. Recommended Dilutions: Western Blot: 0.5-2ug/ml. Detects Nck beta in RIPA lysates of murine testes and spleen homogenates. Mouse testes homogenate was resolved by electrophoresis, transferred to nitrocellulose and probed with N0550-50 (2ug/ml). Proteins were visualized using an IgG (HRP) goat anti-rabbit and a chemiluminescence detection system. Immunoprecipitation: 1:100. Immunoprecipitates Nck beta from 1-2mg of mouse tissue homogenates. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
