Approximately three-quarters of today’s drugs act upon some type of GPCR. A major focus of drug discovery research is finding more effective drugs that act upon these targets, as well as characterizing new ‘orphan’ receptors and validating them as drug targets. With the Thermo Scientific Cellomics GPCR Signaling BioApplication, these types of studies can be streamlined to gain critical information more rapidly than ever before. This BioApplication utilizes three fluorescent probes for image analysis: nuclear, cell membrane, and target molecule (for example GFP-receptor, GFP-barrestin, or fluorescent ligand). The application is able to detect and classify the different sub-cellular distribution phenotypes for fluorescent receptor complexes, such as colocalization with the cell membrane, clathrin-coated pits, and endosomes. The GPCR Signaling BioApplication can distinguish among any of these phenotypes, making it universally applicable to receptors of different classes and types.

The GPCR Signaling BioApplication has been validated with the β2 Adrenergic receptor (see posters). For this analysis, half of each triplicate 96-well microplate was treated with medium only while the other half was treated with a maximal dose of agonist (370 nm isoproterenol). The Z’ factor and other measures of assay performance were then calculated for each plate:

Analysis of these “Min-Max” plates indicated exceptional performance with a Z’ window of 0.73. This value, along with high signal-to-noise (S/N) and signal-to-background (S/B) ratios (Table, left) indicated that the GPCR Signaling BioApplication provides a robust analysis of GPCR activation.