OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
The NanoBRET™ Target Engagement (TE) Intracellular Kinase Assays are based on the Bioluminescence Resonance Energy Transfer (BRET) system, an energy transfer technique designed to measure molecular proximity in living cells. The assay measures the apparent affinity of test compounds by competitive displacement of the NanoBRET™ tracer, reversibly bound to a NanoLuc® luciferase-kinase fusion expressed in cells.
Family:
Kinase
Sub Family:
PtdIns(4,5)P 3-Kinase
Class:
Lipid
Protein Name:
PI3Kalpha
Uniprot Number:
P42336
Protein Aliases:
PI3-kinase subunit alpha|PI3K-alpha PI3Kalpha|PtdIns-3-kinase subunit alpha|Phosphatidylinositol 4,5-bisphosphate 3-kinase 110 kDa catalytic subunit alpha|PtdIns-3-kinase subunit p110-alpha|Phosphoinositide 3-kinase alpha Phosphoinositide-3-kinase catalytic alpha polypeptide|Serine/threonine protein kinase PIK3CA
Gene Name:
PIK3CA
Gene ID:
5290
Gene Aliases:
MCM| CCM4| CWS5| MCAP| PI3K| CLAPO| CLOVE| MCMTC| PI3K-alpha| p110-alpha
Accession Number:
NM_006218
Organism:
Human
Construct Details:
NanoLuc® luciferase fused to the N-terminus of the Wild Type Full-length kinase
Cell Type:
293H
Assay Information
Assay Type:
Functional
Assay Sub Type:
Cell Based Binding
Functional Mode:
Antagonist
Detection Method:
NanoBRET
Testing Information
Procedure Summary:
NanoBRET TE Intracellular Kinase Assays are performed according to Promega Adherent Format Assay Protocol using NanoBRET™ Tracer K-3. 293H cells are transfected with NanoLuc®-PIK3CA Fusion Vector and co-transfected with PIK3R1 Expression Vector, seeded in 384-well plates, and cultivated overnight. Target engagement of compounds are measured by treating the cells with the compound in presence of a fixed concentration of fluorescent tracer.
Tracer:
NanoBRET™ Tracer K-3
Incubation:
120 min at 37°C
Test Sample Requirements:
Minimum 1) Dose Response: 120 µL of 10 mM stock -OR- 1 mg (pre-weighed) for a top concentration at 30 µM)
Turnaround Time
Standard:
25 Business Days
Additional Information
Brand:
LeadHunter
Testing Location:
France - Celle L'Evescault
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