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:

CMGC

Class:

Protein Ser/Thr

Protein Name:

GSK3beta

Uniprot Number:

P49841

Protein Aliases:

GSK-3 beta|Serine/threonine-protein kinase GSK3B|EC 2.7.11.1

Gene Name:

GSK3B

Gene ID:

2932

Accession Number:

NM_001146156

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-8. 293H cells are transfected with NanoLuc®-GSK3B Fusion 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:

K-8 (0.055 µM)

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:

15 Business Days

Reference Compound Data

PubChem ID

57654618

Name

Staurosporine

Measurement

180nM - IC50

PubChem ID

45111335

Name

APY-69

Measurement

1.65nM - IC50

Additional Information

Brand:

LeadHunter

Testing Location:

France - Celle L'Evescault