2 articles published in JoVE
NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases Brian J. Stockman1, Abinash Kaur1, Julia K. Persaud1, Maham Mahmood1, Samantha F. Thuilot1, Melissa B. Emilcar1, Madison Canestrari1, Juliana A. Gonzalez1, Shannon Auletta1, Vital Sapojnikov1, Wagma Caravan1,2, Samantha N. Muellers1,3 1Department of Chemistry, Adelphi University, 2Department of Chemistry, Washington University in St. Louis, 3Department of Chemistry, Boston University NMR-based activity assays have been developed to identify and characterize inhibitors of two nucleoside ribohydrolase enzymes. Protocols are provided for initial compound assays at 500 μM and 250 μM, dose-response assays for determining IC50 values, detergent counter screen assays, jump-dilution counter screen assays, and assays in E. coli whole cells.
Preparation of Mitochondrial Enriched Fractions for Metabolic Analysis in Drosophila Eugenia Villa-Cuesta1, David M. Rand2 1Biology Department, Adelphi University, 2Department of Ecology and Evolutionary Biology, Brown University Mitochondria play central roles in the regulation of metabolism and homeostasis. Subtle changes in mitochondrial metabolism that affect organismal physiology could be difficult to detect in whole organism metabolomics studies. Here we describe an isolation method that enhances the detection of subtle metabolic shifts in Drosophila melanogaster.