Microtubule dynamics : methods and protocols
Author(s)
Bibliographic Information
Microtubule dynamics : methods and protocols
（Methods in molecular biology / John M. Walker, series editor, 777）（Springer protocols）
Humana, c2011
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Includes bibliographical references and index
Description and Table of Contents
Description
Microtubules are at the heart of cellular self-organization, and their dynamic nature allows them to explore the intracellular space and mediate the transport of cargoes from the nucleus to the outer edges of the cell and back. In Microtubule Dynamics: Methods and Protocols, experts in the field provide an up-to-date collection of methods and approaches that are used to investigate microtubule dynamics in vitro and in cells. Beginning with the question of how to analyze microtubule dynamics, the volume continues with detailed descriptions of how to isolate tubulin from different sources and with different posttranslational modifications, methods used to study microtubule dynamics and microtubule interactions in vitro, techniques to investigate the ultrastructure of microtubules and associated proteins, assays to study microtubule nucleation, turnover, and force production in cells, as well as approaches to isolate novel microtubule-associated proteins and their interacting proteins. Written in the highly successful Methods in Molecular Biology (TM) 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.
Definitive and practical, Microtubule Dynamics: Methods and Protocols provides the key protocols needed by novices and experts on how to perform a broad range of well-established and newly-emerging techniques in this vital field.
Table of Contents
1. How to Measure Microtubule Dynamics?
Anne Straube
2. Purification of Tubulin from Porcine Brain
Christopher Gell, Claire T. Friel, Barbara Borgonovo, David N. Drechsel, Anthony A. Hyman, and Jonathon Howard
3. Purification of Tubulin from the Fission Yeast Schizosaccharomyces pombe
Douglas R Drummond, Susan Kain, Anthony Newcombe, Christina Hoey, Miho Katsuki, and Robert A. Cross
4. Generation of Differentially Polyglutamylated Microtubules
Benjamin Lacroix and Carsten Janke
5. Preparation of Pure Tyrosinated or Detyrosinated Tubulin Isoforms
Laurence Lafanechere and Didier Job
6. ab-Tubulin and Microtubule Binding Assays
Jaime N. Campbell and Kevin C. Slep
7. Analysis of Microtubule Assembly Kinetics Using Turbidimetry
Felicia Gaskin
8. Isolation of Centrosomes from Cultured Cells
Simone Reber
9. Preparation of Dual-Color Polarity-Marked Fluorescent Microtubule Seeds
Miho Katsuki, Etsuko Muto, and Rob Cross
10. Reconstitution and Quantification of Dynamic Microtubule End Tracking In Vitro Using TIRF Microscopy
Ivo A. Telley, Peter Bieling, and Thomas Surrey
11. Force Generation by Dynamic Microtubules In Vitro
Svenja-Marei Kalisch, Liedewij Laan, and Marileen Dogterom
12. Variations on the Single Molecule Assay for Microtubule-Associated Proteins and Kinesins
Susanne Bechstedt, Michal Wieczorek, Michael Noujaim, and Gary J. Brouhard
13. Analyzing the ATP Hydrolysis Cycle of Microtubule Motors
Claire T. Friel, Clive R. Bagshaw, and Jonathon Howard
14. Cryo-Electron Tomography of Microtubules Assembled In Vitro from Purified Components
Frederic M. Coquelle, Sophie Blestel, Claire Heichette, Isabelle Arnal, Charles Kervrann, and Denis Chretien
15. Kinetochore Microtubules, Analyzed by Correlated Light and Immuno-Electron Microscopy
Andreas Merdes and Jan De Mey
16. Photoactivatable-GFP-a-Tubulin as a Tool to Study Microtubule Plus-End Turnover in Living Human Cells
Catarina P. Samora and Andrew D. McAinsh
17. Quantification of Asymmetric Microtubule Nucleation at Sub-Cellular Structures
Xiaodong Zhu and Irina Kaverina
18. Investigating Microtubule Dynamic Instability Using Microtubule Targeting Agents
Stephane Honore and Diane Braguer
19. Laser Ablation of the Microtubule Cytoskeleton: Setting Up and Working with an Ablation System
Nicola Maghelli and Iva Tolic-Norrelykke
20. Isolation, Identification, and Validation of Microtubule-Associated Proteins from Drosophila Embryos
Robin Antrobus and James G. Wakefield
21. Isolation of Novel +TIPs and Their Binding Partners Using Affinity Purification Techniques
Ka Lou Yu, Nanda Keijzer, Casper C. Hoogenraad, and Anna Akhmanova
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