Catalog # DP2241

Dok1 (Tyr-362)[Dok2 (Tyr-337)], phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 62 kDa

Doks are a family of adaptor proteins that include six Dok proteins (Dok1 to Dok6), which have an N-terminal pleckstrin homology domain, a central phosphotyrosine binding domain, and a C-terminal region containing multiple tyrosine residues. When phosphorylated, these tyrosines can serve as docking sites for SH2 domain-containing proteins. Dok1 (p62dok) has been shown to bind Ras-GAP, Nck, and Csk. Several tyrosine phosphorylation sites have been identified for Dok1. One site, Tyr-362 (Tyr-361 mouse), is phosphorylated by c-Abl, is required for Nck binding, and may be critical for filopodia formation during fibroblast spreading on fibronectin. Alternatively, Dok1 activity is also regulated by serine phosphorylation. IκB Kinase β phosphorylates several serine sites including Ser-450 in vitro, and TNFα, IL-1, and radiation treatment lead to phosphorylation of Ser-443, Ser-446, and Ser-450 in vivo. Phosphorylation of these serine sites may be required for Dok-mediated inhibition of MAPK signaling and stimulation of cell motility.

References

Lee, S. et al. (2004) Proc. Nat. Acad. Sci. 101(50):17416.
Woodring, P.J. (2004) J Cell Biol. 165(4):493.
Kashige, N. et al. (2000) Proc. Nat. Acad. Sci. 97(5):2093.
Noguchi, T. et al. (1999) EMBOJ 18(7):1748.

Western blot image of Jurkat cells stimulated with calyculin A (100 nM, 30 min) (lanes 1-6) followed by lambda phosphatase (lanes 2 & 6) or alkaline phosphatase (lane 4) treatment. The blots were probed with anti-Dok1 (lanes 1 & 2), anti-Dok1 (Tyr-362) (lanes 3 & 4), and anti-Dok1 (Ser-450) (lanes 5 & 6).

Phospho-Dok1 (Tyr-362) synthetic peptide (coupled to KLH) corresponds to amino acids surrounding tyrosine 362 in human Dok1. This sequence is conserved in Dok1 from rat and mouse (Tyr-361), and has high homology to Dok2 (Tyr-337). The site is not conserved in other Dok family members.

*For more information, see UniProt Accession Q99704

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.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody was cross-adsorbed to a phosphotyrosine peptide before affinity purification using phospho-Dok1 (Tyr-362) peptide. The purified antibody detects a band at 62 kDa* corresponding to Dok1 in western blots of human Jurkat cells, but does not detect this band after alkaline phosphatase treatment. Similar to Dok1 (Ser-450) antibody, this antibody also detects an 80 kDa band that has not been identified.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: