The NanoBRET™ Target Engagement (TE) Intracellular Kinase Assay:
Measures compound binding at select target kinases in intact cells. The assay is based on the NanoBRET™ System, an energy transfer technique designed to measure molecular proximity in living cells. The NanoBRET™ TE Assay measures the apparent affinity of test compounds by competitive displacement of the NanoBRET™ tracer, reversibly bound to a NanoLuc® luciferase-kinase fusions in cells.
How it Works:
Using the NanoBRET™ TE Kinase Assay, a fixed concentration of tracer (K-4 or K-5) is added to cells expressing the desired NanoLuc® kinase fusion to generate a BRET signal. Introduction of competing compounds results in a dose-dependent decrease in NanoBRET™ signal, which allows quantitation of the intracellular affinity of the target protein for the test compound.