FAK Human TK Kinase Enzymatic Radiometric Assay [10 μM ATP] - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
PTK2 (Protein Tyrosine Kinase 2, formerly FAK or Focal Adhesion Associated Protein Tyrosine Kinase) is a widely expressed nonreceptor involved in the structure and function of focal adhesions, which are critical for cellular integrity by maintaining cell-cell and cell-matrix interactions. Integrin-mediated cell adhesion causes activation of MAP kinases and increased tyrosine phosphorylation of PTK2. Autophosphorylation of PTK2 leads to the binding of SH2-domain proteins including Src-family kinases and the Grb2-Sos complex, then activate Ras cascade and ultimately to mitogen activated protein (MAP) kinase activation.
Family:
Kinase
Sub Family:
TK
Class:
Protein Tyrosine
Protein Name:
FAK
Uniprot Number:
Q05397
Protein Aliases:
protein phosphatase 1, regulatory subunit 71
Gene Name:
PTK2
Gene ID:
5747
Gene Aliases:
FAK|FADK|FAK1|PPP1R71
Accession Number:
NM_005607
Organism:
Human
Construct Details:
Fragment (2-1052), 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 PTK2 expressed in insect Sf9 cells is used. Test compound and/or vehicle is preincubated with 2.4 µg/ml enzyme for 15 minutes at 37°C in modified HEPES buffer pH 7.4. The reaction is initiated by addition of 0.2 mg/ml poly(Glu:Tyr), 10 µM ATP containing 0.05 µCi [γ³²P]ATP for 30 minutes and terminated by further addition of 3% H₃PO₄. An aliquot is counted to determine the radioactivity of [³²P]Poly(Glu:Tyr). 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
Adverse / Beneficial Action:
PTK2 (FAK) inhibitors may be beneficial to treat inflammation and various tumors.
Additional Information
Brand:
Panlabs
Testing Location:
Taiwan - Taipei
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