Using Promega’s NanoBRET technology, scientists at Aurelia Bioscience have implemented intracellular kinase assays designed to study the interaction of compounds on kinase targets in intact living cells.
Advantages of this technique include examination of compound binding to target kinases at physiological ATP concentrations, expression of full-length kinase target in cells, disruption of cells (with digitonin) to release cellular content from the cell, and, examination of the association (K-on) and dissociation (K-off) rates of compounds in live cells. It can also be used to look at allosteric inhibitors in addition to ATP competitive inhibitors. This technique is applicable to a wide range of kinases and can be performed as a pseudo biochemical binding assay.
Watch this video to learn more.