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Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications
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Chapters
- 00:05Title
- 01:37Expansion and Cryopreservation of Assay Ready Cells
- 02:58Plating Assay Ready Cells
- 04:15Small Molecule Screening and Persistent Agonist Treatment
- 05:16Stimulation, Quenching, and Estimation of cAMP Accumulation
- 07:02Results: Development and Testing of a Cell-based 384 Well Sensitization Assay with Receptor Antagonists
- 09:12Conclusion
Persistent activation of inhibitory G protein-coupled receptors results in sensitization of adenylyl cyclase signaling. To identify the essential molecular pathways, nonbiased approaches are necessary; however, this strategy requires the development of a scalable cell-based cAMP sensitization assay. Herein, we describe a sensitization assay for small molecule and siRNA screening.
