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Technical Data Sheet

AK6210

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c-Abl 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
AP1271 Skora, L. et al. (2015) Eur J Haematol. doi: 10.1111 ELISA: K562, KCL-22, and Ba/F3 cell lines
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
Kit Summary
The c-Abl phospho-regulation antibody sampler kit can be used to detect c-Abl activation resulting from phosphorylation of Tyr-245 and Tyr-412. The kit also includes an antibody to examine total c-Abl expression levels and secondary reagents for rabbit polyclonal and mouse monoclonal antibody detection.
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Western blot analysis of K-562 cells treated with pervanadate (1 mM) for 30 minutes (lanes 1, 3, & 5). Some lanes were treated with alkaline phosphatase to remove phosphorylation on c-Abl (lanes 2, 4, & 6), then the blots were probed with anti-c-Abl (lanes 1 & 2), anti-c-Abl (Tyr-412) (AP1271; lanes 3 & 4), or anti-c-Abl (Tyr-245) (AP1251; lanes 5 & 6).

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Western blot analysis of K-562 cells treated with pervanadate (1 mM) for 30 minutes. Blots were probed with anti-c-Abl (lane 1), anti-c-Abl (Tyr-412) (lanes 2-5), or anti-c-Abl (Tyr-245) (lanes 6-9). In some lanes, the antibodies were used in the absence (lane 2 & 6) or presence of c-Abl (Tyr-412) (lane 4 & 8) or c-Abl (Tyr-245) (lane 3 & 7) blocking peptides, or BSA conjugated to phospho-tyrosine (lane 5 & 9).

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Background
The c-Abl proto-oncogene encodes a nonreceptor type protein tyrosine kinase that is widely expressed and is distributed in both the nucleus and the cytoplasm of cells. It has been implicated in regulation of cell proliferation, differentiation, apoptosis, cell adhesion, and stress response. A variety of stimuli activate c-Abl kinase including integrin activation, PDGF stimulation, and binding to proteins, such as c-Jun. Tyrosine phosphorylation is important for the regulation of c-Abl kinase activity. Tyrosine 245 is located in the linker region between the SH2 and catalytic domains. Phosphorylation of Tyr-245 is involved in activation of c-Abl kinase activity. Tyrosine 412 is located in the kinase activation loop of c-Abl, and phosphorylation of this residue is required for kinase activity. Thus, phosphorylation of Tyr-245 and Tyr-412 may be critical for activation of c-Abl in a variety of cell signaling pathways.

Background References
Van Etten, R.A. et al. (1999) Trends Cell. Biol. 9:179.
Brasher, B.B. et al. (2000) J. Biol. Chem. 275:35631.
Pluk, H. et al. (2002) Cell 108:247.
Buffer and Storage
Rabbit polyclonal and mouse monoclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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