Straube Microtubule Dynamics

Methods and Protocols
Erscheinungsjahr 2011
ISBN: 978-1-61779-252-6
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 777, 319 Seiten, eBook

Reihe: Methods in Molecular Biology

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™ 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.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Straube, Anne

Inhaltsverzeichnis

How to Measure Microtubule Dynamics?.- Purification of Tubulin from Porcine Brain.- Purification of Tubulin from the Fission Yeast Schizosaccharomyces pombe.- Generation of Differentially Polyglutamylated Microtubules.- Preparation of Pure Tyrosinated or Detyrosinated Tubulin Isoforms.- ab-Tubulin and Microtubule Binding Assays.- Analysis of Microtubule Assembly Kinetics Using Turbidimetry.- Isolation of Centrosomes from Cultured Cells.- Preparation of Dual-Color Polarity-Marked Fluorescent Microtubule Seeds.- Reconstitution and Quantification of Dynamic Microtubule End Tracking In Vitro Using TIRF Microscopy.- Force Generation by Dynamic Microtubules In Vitro.- Variations on the Single Molecule Assay for Microtubule-Associated Proteins and Kinesins.- Analyzing the ATP Hydrolysis Cycle of Microtubule Motors.- Cryo-Electron Tomography of Microtubules Assembled In Vitro from Purified Components.- Kinetochore Microtubules, Analyzed by Correlated Light and Immuno-Electron Microscopy.- Photoactivatable-GFP-a-Tubulin as a Tool to Study Microtubule Plus-End Turnover in Living Human Cells.- Quantification of Asymmetric Microtubule Nucleation at Sub-Cellular Structures.- Investigating Microtubule Dynamic Instability Using Microtubule Targeting Agents.- L Isolation, Identification, and Validation of Microtubule-Associated Proteins from Drosophila Embryos.- Isolation of Novel +TIPs and Their Binding Partners Using Affinity Purification Techniques.