Abstract
Recent developments in optical microscopy and nanometer tracking have greatly improved our understanding of cytoskeletal motor proteins. Using fluorescence microscopy, dynamic interactions are now routinely observed in vitro on the level of single molecules mainly using a geometry, where fluorescently labeled motors move on surface-immobilized filaments. In this chapter, we review recent methods related to single-molecule kinesin motility assays. In particular, we aim to provide practical advice on: how to set up the assays, how to acquire high-precision data from fluorescently labeled kinesin motors and attached quantum dots, and how to analyze data by nanometer tracking.
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Acknowledgments
The authors would like to thank all current and former members of the Howard and Diez labs for the tremendous efforts in helping to develop the described protocols.
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Korten, T., Nitzsche, B., Gell, C., Ruhnow, F., Leduc, C., Diez, S. (2011). Fluorescence Imaging of Single Kinesin Motors on Immobilized Microtubules. In: Peterman, E., Wuite, G. (eds) Single Molecule Analysis. Methods in Molecular Biology, vol 783. Humana Press. https://doi.org/10.1007/978-1-61779-282-3_7
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DOI: https://doi.org/10.1007/978-1-61779-282-3_7
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