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NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases
 

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

Article DOI: 10.3791/59928-v • 10:24 min • June 30th, 2019
June 30th, 2019 •
1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1,2, 1,3
1Department of Chemistry, Adelphi University, 2Department of Chemistry, Washington University in St. Louis, 3Department of Chemistry, Boston University

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Resumo
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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.

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Keywords: NMR-based Activity Assays Enzyme Inhibition IC50 Values Artifact Detection Whole-cell Activity Nucleoside Ribohydrolases Fragment Screening Trichomonas Vaginalis Anti-trichomonal Drugs Reagent Volumes Reaction Components Substrate Preparation Test Compound Preparation Reaction Stock Solution Reaction Initiation Reaction Quenching
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