This chapter describes methods for high-speed, unloaded, in vitro single-molecule kinesin tracking experiments. Instructions are presented for constructing a total internal reflection dark-field microscope (TIRDFM) and labeling motors with gold nanoparticles. An AMP-PNP unlocking assay is introduced as a specialized means of capturing processive events in a reduced field of view. Finally, step-finding tools for analyzing high frame-rate tracking data are described.
CITATION STYLE
Mickolajczyk, K. J., & Hancock, W. O. (2018). High-resolution single-molecule kinesin assays at kHz frame rates. In Methods in Molecular Biology (Vol. 1805, pp. 123–138). Humana Press Inc. https://doi.org/10.1007/978-1-4939-8556-2_7
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