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-g. Nck is an oncogenic protein and 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. Nck binds through its second SH3 domain to WIP, and Nck binds to WIP at a site (aa321-415).

Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.

Western Blot: 0.1-1ug/ml detected Nck in human A431 carcinoma cells. Previous lots detected Nck in mouse 3T3/A31 fibroblasts and rat L6 skeletal fibroblasts. Non-stimulated A431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with N0550-05 (0.1ug/ml). Proteins were visualized using IgG (HRP) goat anti-rabbit and a chemilluminescence detection system. Immunoprecipitation: 5ug immunprecipitates Nck from a lysate of human A431 carcinoma cells, mouse 3T3/A31 fibroblasts and rat L6 skeletal fibroblasts. Optimal dilutions to be determined by the researcher.

A0001-50: A431 Cells, Non-Stimulated in RIPA Buffer, Sample Buffer.

Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: MaxLight™650 conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Note: Applications are based on unconjugated antibody.

Purified GST fusion protein corresponding to aa1-254 of human Nck, produced in E. coli.

Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with MaxLight™650.

Purified by Protein G affinity chromatography.

Recognizes human Nck at ~47kD. Species Crossreactivity: mouse and rat