New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories. (March 2016)

• Record Type: Journal Article
• Authors: Tewson, Paul H.; Martinka, Scott; Shaner, Nathan C.; Hughes, Thomas E.; Quinn, Anne Marie
• Is Part Of: Journal of biomolecular screening. Volume 21:Number 3(2016)
• Page Start: 298
• Page End: 305
• Publication Date: 2016-03
• Subjects: fluorescence -- baculovirus -- mNeonGreen -- GFP -- Z′ statistic -- live-cell assay -- adenylyl cyclase -- cAMP -- diacyl glycerol -- DAG; Drugs -- Analysis -- Periodicals; Drugs -- Testing -- Periodicals; Biomolecules -- Analysis -- Periodicals; 572.36
• DOI: 10.1177/1087057115618608
• Languages: English
• ISSNs: 1087-0571

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.