Western blot analysis of extracts of H526 cells, untreated or treated with SCF, using Phospho-c-Kit (Tyr703) (D12E12) Rabbit mAb (upper) or c-Kit Antibody #3392 (lower).
Western blot analysis of extracts from MO7e cells, untreated or stimulated with SCF #9907 (50 ng/ml for 2 minutes), using Phospho-c-Kit (Tyr719) Antibody (upper) or c-Kit Antibody #3392 (lower).
Western blot analysis of extracts from NCI-H526 cells using c-Kit (D13A2) XP® Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Confocal immunofluorescent analysis of NCI-H526 (left) and Jurkat (right) cells using c-Kit (D13A2) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
|Phospho-c-Kit (Tyr703) (D12E12) Rabbit mAb 3073||20 µl||
|Phospho-c-Kit (Tyr719) Antibody 3391||20 µl||
||H M||120, 145||Rabbit|
|c-Kit (D13A2) XP® Rabbit mAb 3074||20 µl||
||H M||120 and 145||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The c-Kit Antibody Sampler Kit provides a fast and economical means of evaluating levels of c-Kit receptor protein phosphorylated at the specified sites, as well as total c-Kit receptor levels. The kit contains enough primary and secondary antibody to perform two Western blot experiments.
Phospho-c-Kit antibodies detect endogenous levels of c-Kit when phosphorylated at the specified sites. c-Kit (D13A2) XP® Rabbit mAb detects endogenous levels of total c-kit receptor protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the residues surrounding Tyr703 of human c-Kit. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr719 of mouse c-Kit. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
c-Kit is a member of the subfamily of receptor tyrosine kinases that includes PDGF, CSF-1, and FLT3/flk-2 receptors (1,2). It plays a critical role in activation and growth in a number of cell types including hematopoietic stem cells, mast cells, melanocytes, and germ cells (3). Upon binding with its stem cell factor (SCF) ligand, c-Kit undergoes dimerization/oligomerization and autophosphorylation. Activation of c-Kit results in the recruitment and tyrosine phosphorylation of downstream SH2-containing signaling components including PLCγ, the p85 subunit of PI3 kinase, SHP2, and CrkL (4). Molecular lesions that impair the kinase activity of c-Kit are associated with a variety of developmental disorders (5), and mutations that constitutively activate c-Kit can lead to pathogenesis of mastocytosis and gastrointestinal stromal tumors (6). Tyr719 is located in the kinase insert region of the catalytic domain. c-Kit phosphorylated at Tyr719 binds to the p85 subunit of PI3 kinase in vitro and in vivo (7).
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