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

PubChem ID

5328760

Name

Tyrphostin AG 538

Measurement

0.04μM - IC50

PubChem ID

122130844

Name

Staurosporine

Measurement

0.0064μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei