NanoBRET™ Assay for Monitoring of Ligand Binding to GPCRs in Live Cells, Using the CLARIOstar® and the PHERAstar FS

Due to the crucial role of G protein-coupled receptors (GPCRs) in mediating cellular responses to external stimuli, these receptors have been and will remain a prime focus for medical research and the pharmaceutical industry for many years to come. Their importance is especially highlighted by the statistic that 30-50% of marketed pharmaceuticals target GPCRs. BRET (bioluminescence resonance energy transfer) requires only the addition of a suitable substrate to generate donor light emission, as well as energy that can excite fluorescently-labelled ligands in a non-radiative manner. The donor luciferase enzyme can be expressed on the N-terminus of the receptor of interest in an appropriate transfected cell line. This application note demonstrates the first use of BRET for monitoring ligand binding to GPCRs in live cells enabled by NanoLuc® luciferase.