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Updated: Jun 4, 2025

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays
Published on: February 4, 2021
Expression, Purification, and In Vitro Analysis of Myosin
Glenn Carrington1,2, Yasuharu Takagi3,4, Oliver Umney1,2,4
1Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, UK.
Abstract:
To understand the mechanics and kinetic properties of cytoskeletal molecular motors such as myosin, typically the motor of interest needs to be expressed and purified and then analyzed using a range of in vitro-based assays. In this chapter, we describe how to express and purify myosin using the insect cell system, how to characterize the purified protein by mass photometry and negative-stain EM to assess its quality, and how to perform in vitro assays in which fluorescently labeled myosin walks along actin tracks, including a brief description of adapting these assays for MINFLUX imaging.
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