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

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

DOI:
10.3791/62180-v

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08:57 min

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February 04, 2021

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, , , ,
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

Chapters

  • 00:05Introduction
  • 01:04Gliding Actin Filament Assay
  • 05:02Performing the Myosin 5a TIRF Microscopy Assay
  • 06:40Results: Gliding Activity Assay of the NM2b and M5a-HMM Filaments
  • 08:05Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

MyosinIn VitroFluorescence MicroscopyMotility AssaysNon-muscle MyosinProtein DynamicsSingle MoleculeEnsembleCytoskeletal SystemsKinesinsDyneinsMicrotubulesNitrocelluloseFlow Chamber

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