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This volume details comprehensive state-of-the-art methods on actin microfilaments and microtubules and how they work to achieve different cellular functions in different cellular contexts. Chapters guide readers through protein purification, in vitro reconstitution of several cytoskeleton properties, analyses of microtubule- and actin-based structures, functional dissection of post-translational modifications, and roles in several biological processes. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Cytoskeleton Dynamics: Methods and Protocols aims to provide a wide range of experimental approaches and be an invaluable resource for present and future generations of cytoskeleton researchers.
Purification of Mammalian Tubulins and Tubulin-Associated Proteins Using a P2A-Based Expression System
Expression and Purification of Microtubule-Associated Proteins from HEK293T Cells for In Vitro Reconstitution
In Vitro Reconstitution Assays of Microtubule Amplification and Lattice Repair by the Microtubule-Severing Enzymes Katanin and Spastin
In Vitro Microtubule Dynamics Assays Using Dark-Field Microscopy
In Vitro Reconstitution of Dynamic Co-organization of Microtubules and Actin Filaments in Emulsion Droplets
Studying Tau-Microtubule Interaction Using Single-Molecule TIRF Microscopy
In Vitro FRET- and Fluorescence-Based Assays to Study Protein Conformation and Protein-Protein Interactions in Mitosis
Methods for Studying Membrane-Associated Bacterial Cytoskeleton Proteins In Vivo by TIRF Microscopy
Processing TIRF Microscopy Images to Characterize the Dynamics and Morphology of Bacterial Actin-Like Assemblies
Studying Centriole Duplication and Elongation in Human Cells
Assaying Microtubule Nucleation
Detection of Microtubule Nucleation Hotspots at the Golgi
Ultrastructural Analysis of Microtubule Ends
Optogenetic Control of Microtubule Dynamics
Measurement of Microtubule Half-Life and Poleward Flux in the Mitotic Spindle by Photoactivation of Fluorescent Tubulin
Functional Dissection of Mitosis Using Immortalized Fibroblasts from the Indian Muntjac, a Placental Mammal with Only Three Chromosomes
Visualization and Functional Analysis of Spindle Actin and Chromosome Segregation in Mammalian Oocytes
Using the Four-CellC. elegans Embryo to Study Contractile Ring Dynamics During Cytokinesis
Knocking Out Multiple Genes in Cultured Primary Neurons to Study Tubulin Posttranslationa​l Modifications
Measuring the Impact of Tubulin Posttranslationa​l Modifications on Axonal Transport
Methods to Measure Perinuclear Actin Dynamics During Nuclear Movement in Migrating Cells