Biological Information
Background Information:
Binding of stem cell factor (SCF) to C-Kit receptor tyrosine kinase induces homodimerization and intermolecular tyrosine phosphorylation of the receptor, creating docking sites for a number of SH2-containing signal transduction molecules. These include phosphatidylinositol 3'-kinase (PI 3K) and phospholipase C-1, Stat1, the Src family kinases Src, Lyn, and Fyn, and the GTPase activating protein GAP. Mutation in Kit which result in ligand-independent activation of the kinase were found to be associated with human gastrointestinal stromal tumors and mastocytosis.
Family:
Kinase
Sub Family:
RTK
Class:
Protein Tyrosine
Protein Name:
Kit
Uniprot Number:
P10721
Protein Aliases:
Piebald trait protein|PBT|Proto-oncogene c-Kit|Tyrosine-protein kinase Kit|p145 c-kit|v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog|CD antigen CD117
Gene Name:
KIT
Gene ID:
3815
Gene Aliases:
CD117|SCFR|C-Kit
Accession Number:
NM_000222
Organism:
Human
Construct Details:
Fragment (544-976), Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometric
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human recombinant protein kinase C-Kit expressed in insect High-5 cells is used. Test compound and/or vehicle is preincubated with 3.3 µg/ml enzyme‡ in modified HEPES buffer pH 7.4 for 15 minutes at 37°C. The reaction is initiated by addition of 200 µg/ml poly(Glu:Tyr), 10 µM ATP and 0.05 µCi [γ³²P]ATP for another 30 minute incubation period and terminated by further addition of 3% H3PO4. An aliquot is removed and counted to determine the amount of [³²P]Poly(Glu:Tyr) formed. Compounds are screened at 10 µM.
Substrate:
200 µg/ml Poly(Glu:Tyr)
ATP Concentration:
10 uM
Incubation:
30 min at 37°C
Control Inhibitor:
Staurosporine
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
10
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
Panlabs
Testing Location:
Taiwan - Taipei