title: New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories.
abstract: Protein-based, fluorescent biosensors power basic research on cell signaling in health and disease, but their use in automated laboratories is limited. We have now created two live-cell assays, one for diacyl glycerol and another for cAMP, that are robust (Z' > 0.7) and easily deployed on standard fluorescence plate readers. We describe the development of these assays, focusing on the parameters that were critical for optimization, in the hopes that the lessons learned can be generalized to the development of new biosensor-based assays.
authors.affiliation: Montana Molecular, Bozeman, MT, USA.
authors.forename: Paul H
authors.initials: PH
authors.lastname: Tewson
authors.affiliation: Montana Molecular, Bozeman, MT, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Martinka
authors.affiliation: Scintillon Institute, San Diego, CA, USA.
authors.forename: Nathan C
authors.initials: NC
authors.lastname: Shaner
authors.affiliation: Montana Molecular, Bozeman, MT, USA.
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Hughes
authors.affiliation: Montana Molecular, Bozeman, MT, USA amq@montanamolecular.com.
authors.forename: Anne Marie
authors.initials: AM
authors.lastname: Quinn
mesh_terms: D057205:Automation, Laboratory; D016367:Baculoviridae / Q000502:physiology; D015374:Biosensing Techniques; D000242:Cyclic AMP / Q000737:chemistry / Q000378:metabolism*; D004075:Diglycerides / Q000737:chemistry / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D015203:Reproducibility of Results; D014161:Transduction, Genetic
chemical_list: D004075:Diglycerides; D000242:Cyclic AMP
keywords: DAG; GFP; Z′ statistic; adenylyl cyclase; baculovirus; cAMP; diacyl glycerol; fluorescence; live-cell assay; mNeonGreen