Agonist and antagonist TRUPATH assays for G protein-coupled receptors
Jeffrey F. DiBerto, Katie Smart, Reid H. J. Olsen, Bryan L. Roth
Abstract
TRUPATH is a bioluminescence resonance energy transfer-based platform for quantifying G protein-coupled receptor activity via dissociation of heterotrimeric G protein biosensors. Here, we present protocols for agonist and antagonist TRUPATH assays in the 384-well plate format, thereby providing an opportunity for higher throughput. We also provide both data analysis and quality control analyses for these assays, along with considerations for assay optimization and solutions for troubleshooting needs that may be encountered. For complete details on the use and execution of this protocol, please refer to Olsen et al. (2020).
Topics & Concepts
Heterotrimeric G proteinG protein-coupled receptorAgonistTroubleshootingAntagonistFörster resonance energy transferReceptorChemistryComputer scienceG proteinPharmacologyBiologyBiochemistryPhysicsFluorescenceQuantum mechanicsOperating systemReceptor Mechanisms and SignalingNeurobiology and Insect Physiology ResearchMass Spectrometry Techniques and Applications