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

PubChem ID

5328760

Name

Tyrphostin AG 538

Measurement

0.45μM - IC50

PubChem ID

122130844

Name

Staurosporine

Measurement

0.073μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

Panlabs

Testing Location:

Taiwan - Taipei