c-Kit is a member of the subfamily of receptor tyrosine kinases that includes PDGF, CSF-1 and FLT3/flk-2 receptors. It plays critical controlling roles in a number of cell types such as hematopoietic stem cells, mast cells, melanocytes and germ cells. Upon binding with its ligand, stem cell factor (SCF), 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 PLCg, the p85 subunit of PI3 kinase, SHP2 and CrkL. Molecular lesions that impair the kinase activity of c-Kit are associated with a variety of developmental disorders, while mutations that constitutively activate c-Kit can lead to pathogenesis of mastocytosis and gastrointestinal stromal tumors. 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. The V560G mutation has been found in patients with gastrointestinal stromal tumors and has been linked to poor prognosis. This mutation confers increased susceptibility to Gleevec®.
Source:Recombinant human cKit (V560G), aa544-end, with N-terminal His tag, expressed in an Baculovirus infected Sf9 cell expression system.
Molecular Weight: 50kD
Specific Activity:115pmol/min/mg
Application:Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling. Other applications not tested.
Reaction Buffer: 50mM HEPES, pH7.5, 10mM MgCl2, 1mM EGTA, 200uM ATP, 0.01% Brij-35 and 2uM substrate (Tyr Peptide 6, Invitrogen) at room temperature for 1 hour.
Storage and Stability:Aliquot to avoid repeated freezing and thawing and store at -70°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for at least 6 months.
