TRKA Human RTK Kinase Enzymatic Radiometric LeadHunter Assay [10 μM ATP] - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Three tyrosine kinase receptors, designated TRKA, TRKB and TRKC, have been identified that selectively bind NGF, BDNF and NT-3, respectively; NT-4/5 also binds to the TRKB receptor. NGF binds to their specific TRK receptor to stimulate intracellular signaling pathways leading to specific protein phosphorylations and differential gene inductions. TRKA (High affinity nerve growth factor, TRK1 transforming tyrosine kinase protein, p140-TrkA, Trk-A) receptor kinases engage the activation of a branched proto-oncoprotein signaling pathway involving Shc, Src, Ras, Raf, mitogen-activated protein kinase, and other downstream effectors. Inhibitors of TRKA may be useful in the treatment of cancer.
Family:
Kinase
Sub Family:
RTK
Class:
Protein Tyrosine
Protein Name:
TRKA
Uniprot Number:
P04629
Protein Aliases:
TRK1-transforming tyrosine kinase protein|Tropomyosin-related kinase A|Tyrosine kinase receptor|Tyrosine kinase receptor A|Trk-A|gp140trk|p140-TrkA|high affinity nerve growth factor receptor
Gene Name:
NTRK1
Gene ID:
4914
Gene Aliases:
TRK|TRKA|MTC
Accession Number:
NM_002529
Organism:
Human
Construct Details:
Fragment (440-end), 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 TRKA expressed in insect Sf21 cells is used. Test compound and/or vehicle is preincubated with 0.18 U/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 0.25 µCi [γ32P]ATP for 30 minutes and terminated by further addition of 3% H3PO4. An aliquot is counted to determine the radioactivity of [32P]P oly(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:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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