Microtubule architecture depends on a complex network of microtubule-associated proteins (MAPs) that act in concert to modulate microtubule assembly/disassembly and spatial arrangement. In vitro reconstitution of cytoskeleton dynamics coupled to single-molecule fluorescence assays has opened new perspectives to quantify the interaction of MAPs with microtubules. Here, we present a Total Internal Reflection Fluorescence (TIRF) microscopy-based assay enabling the characterization of Tau interaction with dynamic microtubules at the single-molecule level. We describe protein sample preparation in flow cells, single-molecule acquisitions by TIRF microscopy, and quantitative analysis of Tau oligomerization states and dwell time on microtubules.
CITATION STYLE
Stoppin-Mellet, V., Bagdadi, N., Saoudi, Y., & Arnal, I. (2020). Studying tau-microtubule interaction using single-molecule TIRF microscopy. In Methods in Molecular Biology (Vol. 2101, pp. 77–91). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0219-5_6
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