Biological Information
Background Information:
BTK (Bruton’s Tyrosine Kinase) is a member of the Tec family kinases which are important components of both antigen receptor signaling and other cell surface receptors, resulting in regulation of the actin cytoskeleton, adhesion, migration, and transcriptional activation. BTK is found in all cells of the hematopoietic lineage except plasma and T cells, and is required for normal B cell development and signal transduction through cell surface molecules. Mutations in the btk gene result in aberrant B cell development, leading to the X-linked agammaglobulinemia phenotype in humans and to the less severe X-linked immunodeficiency in mice. BTK inhibitors may be of benefit in the treatment of leukemias and lymphomas.
Family:
Kinase
Sub Family:
TK
Class:
Protein Tyrosine
Protein Name:
BTK
Uniprot Number:
Q06187
Protein Aliases:
Bruton's tyrosine kinase
Gene Name:
BTK
Gene ID:
695
Gene Aliases:
ATK|XLA|PSCTK1
Accession Number:
NM_000061
Organism:
Human
Construct Details:
Full Length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human recombinant protein kinase BTK expressed in insect Sf21 cells is used. Test compound and/or vehicle is preincubated with 200 ng/ml enzyme‡ in modified HEPES buffer pH 7.4 for 15 minutes at 37°C. The reaction is initiated by addition of 0.2 mg/ml poly(Glu:Tyr), 10 μM ATP and 0.25 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:
LeadHunter
Testing Location:
Taiwan - Taipei