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Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Article DOI: 10.3791/62180-v • 08:57 min • February 4th, 2021
February 4th, 2021 •
1, 1,2, 1, 1, 1
1Laboratory of Molecular Physiology, Cell and Developmental Biology Center, National Heart, Lung and Blood Institute, National Institutes of Health, 2Department of Physiology, Perelman School of Medicine, University of Pennsylvania

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Presented here is a procedure to express and purify myosin 5a followed by a discussion of its characterization, using both ensemble and single molecule in vitro fluorescence microscopy-based assays, and how these methods can be modified for the characterization of nonmuscle myosin 2b.

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Myosin-specific Adaptations In Vitro Fluorescence Microscopy Motility Assays Non-muscle Myosin Dynamics Protein Level Motile Properties Regulatory Conditions Myosin Motility Cellular Behaviors Disease-causing Mutations Single Molecule Ensemble Level Cytoskeletal Systems Purified Kinesins Dyneins Microtubules Fluorophores Chemical Conditions Flow Chambers Nitrocellulose Solution Amyl Acetate Circular Filter Paper Tissue Culture Plate Cover Slips
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