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          Institute: MPI für molekulare Zellbiologie und Genetik     Collection: Publikationen MPI-CBG 2010-arch     Display Documents



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ID: 546680.0, MPI für molekulare Zellbiologie und Genetik / Publikationen MPI-CBG 2010-arch
Microtubule dynamics reconstituted in vitro and imaged by single-molecule fluorescence microscopy
Authors:Gell, Christopher; Bormuth, Volker; Brouhard, Gary J.; Cohen, Daniel N.; Diez, Stefan; Friel, Claire; Helenius, Jonne H.; Nitzsche, Bert; Petzold, Heike; Ribbe, Jan; Schäffer, Erik; Stear, Jeffrey H.; Trushko, Anastasiya; Varga, Vladimir; Widlund, Per; Zanic, Marija; Howard, Jonathon
Place of Publication:Amsterdam, Netherlands
Publisher:Elsevier
Date of Publication (YYYY-MM-DD):2010
Title of Book:Microtubules, in vitro
Start Page:221
End Page:245
Sequence Number of Inbook:13
Full Name of Book-Editor(s):Wilson, Leslie
Copyright:not available
Review Status:not specified
Audience:Experts Only
Intended Educational Use:No
Abstract / Description:In vitro assays that reconstitute the dynamic behavior of microtubules provide insight into the roles of microtubule-associated proteins (MAPs) in regulating the growth, shrinkage, and catastrophe of microtubules. The use of total internal reflection fluorescence microscopy with fluorescently labeled tubulin and MAPs has allowed us to study microtubule dynamics at the resolution of single molecules. In this chapter we present a practical overview of how these assays are performed in our laboratory: fluorescent labeling methods, strategies to prolong the time to photo-bleaching, preparation of stabilized microtubules, flow-cells, microtubule immobilization, and finally an overview of the workflow that we follow when performing the experiments. At all stages, we focus on practical tips and highlight potential stumbling blocks.
External Publication Status:published
Document Type:InBook
Communicated by:nn
Affiliations:MPI für molekulare Zellbiologie und Genetik
Identifiers:LOCALID:4129
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