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

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

English

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9,145 Views

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10:28 min

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August 17, 2019

DOI:

10.3791/60017-v

DOI:
10.3791/60017-v

•

10:28 min
August 17, 2019

•

13 Views
, , , ,
1Department of Molecular Medicine,The Scripps Research Institute, 2Department of Neuroscience,The Scripps Research Institute, 3Laboratory of Molecular Physiology, NHLBI,National Institutes of Health

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.

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Cite this Article

Radnai, L., Stremel, R. F., Sellers, J. R., Rumbaugh, G., Miller, C. A. A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors. J. Vis. Exp. (150), e60017, doi:10.3791/60017 (2019).

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