2 articles published in JoVE
Cycloheximide Chase Analysis of Protein Degradation in Saccharomyces cerevisiae Bryce W. Buchanan1, Michael E. Lloyd1,2, Sarah M. Engle1, Eric M. Rubenstein1 1Department of Biology, Ball State University, 2Bioproduct Research & Development, Eli Lilly and Company Protein abundance reflects the rates of both protein synthesis and protein degradation. This article describes the use of cycloheximide chase followed by western blotting to analyze protein degradation in the model unicellular eukaryote, Saccharomyces cerevisiae (budding yeast).
Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications Jason M. Conley1, Tarsis F. Brust1, Ruqiang Xu1, Kevin D. Burris2, Val J. Watts1 1Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, 2Quantitative Biology, Eli Lilly and Company 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.