OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Eph (erythropoietin-producing hepatocellular) family tyrosine kinase receptors consist of two groups: EphA group members respond to ephrin-A, which is anchored to the cell membrane by glycosylphosphatidylinositol, whereas EphB group members respond to ephrin-B, containing a transmembrane domain. High levels of EphA2 have been documented in many cell models and clinical specimens of human cancer, including metastatic melanoma and breast, prostate, lung, renal cell, colon, and esophageal carcinomas.
Family:
Kinase
Sub Family:
RTK
Class:
Protein Tyrosine
Protein Name:
EphA2
Uniprot Number:
P29317
Protein Aliases:
Epithelial cell kinase|Tyrosine-protein kinase receptor ECK
Gene Name:
EPHA2
Gene ID:
1969
Gene Aliases:
ECK
Accession Number:
NM_004431
Organism:
Human
Construct Details:
596-900 recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometric
Measured Response:
Scintillation
Testing Information
Procedure Summary:
EphA2 (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.1 mg/mL poly(Glu, Tyr) 4:1, 10 mM MgAcetate and [gamma-33P]-ATP (specific activity and concentration as required). The reaction is initiated by the addition of the Mg/ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting.
Substrate:
0.1 mg/mL poly (Glu,Tyr)
Tracer:
33P
ATP Concentration:
[ATP]= 10µM (Km= 158µM )
Incubation:
40 min at Room temperature
Control Inhibitor:
Staurosporine
Test Sample Requirements:
Please provide compounds dissolved in 100% DMSO (PBS or HBSS prohibited).
Stock solution should be ≥50× the final assay concentration (a 10 mM stock is also acceptable).
Required volume:
– <60 kinase assays: 120 µL of 50× stock
– ≥60 assays: 420 µL of 50× stock
If you supply 10 mM stocks or powder (pre-weighed), we will apply the appropriate conversion.
For batches of ~50 compounds supplied as solids, the TAT will be extended.
Minimum Order Quantity:
2
Turnaround Time
Standard:
6 Business Days
Service Scheduling Note:
Assay is run on a weekly schedule beginning on Friday. Compounds and purchase order must be received on or before 13:00 CET Thursday for the order to be included on the service run that begins on Friday of the same week.
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Adverse / Beneficial Action:
Inhibition of EphA2 may be useful in the treatment of various tumors (breast, prostate, lung, colon, ovarian), tumor angiogenesis and metastatic progression.
Additional Information
Brand:
LeadHunter
Testing Location:
KP - France - Celle L'Evescault
RELATED SOLUTIONS
EphA2 Human RTK Kinase Enzymatic Radiometric [Km ATP] KinaseProfiler LeadHunter Assay - FR
Item: 14-560KP
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