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JoVE Journal Biochemistry
A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
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A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors

Article DOI: 10.3791/60017-v • 10:28 min • August 17th, 2019
August 17th, 2019 •
1,2, 1,2, 3, 2, 1,2
1Department of Molecular Medicine, The Scripps Research Institute, 2Department of Neuroscience, The Scripps Research Institute, 3Laboratory of Molecular Physiology, NHLBI, National Institutes of Health

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Summary
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A nicotinamide adenine dinucleotide (NADH)-coupled ATPase assay has been adapted to semihigh throughput screening of small molecule myosin inhibitors. This kinetic assay is run in a 384-well microplate format with total reaction volumes of only 20 µL per well. The platform should be applicable to virtually any ADP producing enzyme.

Tags

NADH-coupled ATPase Assay Screening Small Molecule Inhibitors ATPase Inhibitors Semi-high-throughput Screening Non-muscle Myosin II Inhibitors Methamphetamine Use Disorder Enzymes Producing ATP Actin Solution Microplate Actin Buffer Viscosity Heterogeneity Precipitated Protein Master Enzyme Mix Skeletal Muscle Myosin II LDH Solution PK
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