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).
Bulk Order Anti-Dok1 (Tyr-362) [Dok2 (Tyr-337)], Phosphospecific Antibody
Dok1 (Tyr-362)[Dok2 (Tyr-337)]
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.
Antigen Affinity Purified
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.
Human, Mouse, Rat
Storage at -20°C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20°C.
PBS + 1 mg/ml BSA, 0.05% NaN3 and 50% glycerol
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.
Western blots performed on each lot.
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After date of receipt, stable for at least 1 year at -20°C.