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Kinase Target Engagement

The NanoBRET™ Target Engagement (TE) Intracellular Kinase 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 cellular affinity of test compounds by competitive displacement of a cell-permeable fluorescent NanoBRET™ tracer, reversibly bound to a NanoLuc® luciferase-kinase fusions in cells. 

Promega NanoBRET™ TE kinase assays and specific kinase-NanoLuc® luciferase fusion vectors are used together to measure kinase-compound affinity, occupancy and residence time in live cells. Over 100  kinase specific assays are ready-to-use with representative data available. The assays use a simple multi-well plate format, are scalable, and produce quantitative results.

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Introduction to Target Engagement Assays

Establishing that a small molecule binds with an intended target protein inside live cells forms the basis of target engagement assays. It is an important step in drug discovery as it can enable correlation with compound functional efficacy and can help determine the compound mode of action. In addition, target engagement assays play a central role in the development of highly specific chemical probes that serve as valuable research tools, which can help reveal a target protein’s role in a complex biological system. 

Many existing methods to monitor protein-small molecule binding interactions use purified proteins in biochemical assays, but The results from these biochemical assays may not accurately reflect the binding of the protein-small molecule in living cells. As a result, there is a growing need for quantitative cellular target engagement methods. 

It has been noted in the literature that cellular target engagement methods of high value can (1) quantitate the direct binding between small molecule and target protein; (2) quantitate occupancy or extent of binding of the small molecule to the target; and (3) use live cells, rather than lysates, for these measurements. Promega NanoBRET™ Target Engagement technology possesses these valued attributes.