Figure 7. Representative analysis for binding kinetics of NanoBRET GPCR Tracer G-3 to transiently expressed HiBiT-β₂-AR.

HEK293 cells expressing HiBiT-β₂-AR were assayed as indicated in the protocol. Briefly, cells were transfected, grown overnight, and next day pre-treated with HiBiT detection reagent for 15 min prior to treatment with a titration of NanoBRET GPCR Tracer G-3 (i.e., the previously described propranolol-BODIPY ( Boursier et al., 2020 )). Kinetic measurements of filtered luminescence were taken using GloMax Discover Microplate Reader (Promega) equipped with a 450 nm (8-nm bandpass) filter (donor) and a 600-nm long pass filter (acceptor) (n = 3; Error bars indicate SD).