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Microtubules, in vitro

2nd Edition - August 20, 2013

Editors: John J. Correia, Leslie Wilson

Hardback ISBN:

9 7 8 - 0 - 1 2 - 4 0 7 7 5 7 - 7

eBook ISBN:

9 7 8 - 0 - 1 2 - 4 0 7 8 8 8 - 8

There continues to be intense interest in the microtubule cytoskeleton; the assembly, structure and regulation of microtubules; and the numerous motors and accessory proteins that… Read more

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There continues to be intense interest in the microtubule cytoskeleton; the assembly, structure and regulation of microtubules; and the numerous motors and accessory proteins that control cell cycle, dynamics, organization and transport. The field continues to grow and explore new aspects of these issues driven immensely by developments in optical imaging and tracking techniques. This Second Edition brings together current research and protocols in the field of microtubules in vitro and will serve as a valuable tool for cell biologists, biophysicists and pharmacologists who study the microtubule cytoskeleton, as well as for researchers in the biomedical and biotechnology communities with interest in developing drugs that target microtubules, MAPS and motors.

Series Page
Contributors
Preface

1. Site-Specific Fluorescent Labeling of Tubulin
Abstract
Introduction
Acknowledgments
References

2. Measuring Microtubule Persistence Length Using a Microtubule Gliding Assay
Abstract
Introduction
Summary
Acknowledgments
References

3. Structural Studies of the Doublecortin Family of MAPs
Abstract
Introduction
Acknowledgments
References

4. Detection and Quantification of Microtubule Detachment from Centrosomes and Spindle Poles
Abstract
Introduction
Summary
Acknowledgments
References

5. Regulation of Tubulin Expression by Micro-RNAs: Implications for Drug Resistance
Abstract
Introduction and Rationale
Summary
References

6. Determining the Structure–Mechanics Relationships of Dense Microtubule Networks with Confocal Microscopy and Magnetic Tweezers-Based Microrheology
Abstract
Introduction
Summary
Acknowledgments
References

7. Studying Mitochondria and Microtubule Localization and Dynamics in Standardized Cell Shapes
Abstract
Introduction
Conclusion
Acknowledgments
References

8. Going Solo: Measuring the Motions of Microtubules with an In Vitro Assay for TIRF Microscopy
Abstract
Introduction
Concluding Remarks
Acknowledgments
References

9. Analysis of Microtubules in Isolated Axoplasm from the Squid Giant Axon
Abstract
Preparation of Axoplasm
Analysis of Axoplasmic Microtubule Dynamics
Biochemistry of Axoplasmic Microtubules
Immunohistochemistry of Axoplasmic Microtubules
References

10. Imaging GTP-Bound Tubulin: From Cellular to In Vitro Assembled Microtubules
Abstract
Introduction
Acknowledgments
References

11. Tubulin-Specific Chaperones: Components of a Molecular Machine That Assembles the α/β Heterodimer
Abstract
Introduction
References

12. Heterotrimeric G Proteins and Microtubules
Abstract
Introduction
Concluding Comments
Acknowledgments
References

13. Purification and Biophysical Analysis of Microtubule-Severing Enzymes In Vitro
Abstract
Introduction
Discussion and Summary
Acknowledgments
References

14. Measurement of In Vitro Microtubule Polymerization by Turbidity and Fluorescence
Abstract
Background and Theory
Materials and Equipment
Methods
References

15. Live-Cell Imaging of Microtubules and Microtubule-Associated Proteins in Arabidopsis thaliana
Abstract
Introduction
Concluding Comments
Acknowledgment
References

16. Investigating Tubulin Posttranslational Modifications with Specific Antibodies
Abstract
Introduction
Concluding Remarks
Acknowledgments
References

17. Purification and Assembly of Bacterial Tubulin BtubA/B and Constructs Bearing Eukaryotic Tubulin Sequences
Abstract
Introduction and Rationale
Conclusions
Acknowledgments
References

18. Microtubule-Associated Proteins and Tubulin Interaction by Isothermal Titration Calorimetry
Abstract
Introduction
Conclusion
Acknowledgment
References

19. Methods for Studying Microtubule Binding Site Interactions: Zampanolide as a Covalent Binding Agent
Abstract
Introduction
Summary
Acknowledgments
References

20. Studying Kinetochore-Fiber Ultrastructure Using Correlative Light-Electron Microscopy
Abstract
Introduction
Acknowledgments
References

21. Fluorescence-Based Assays for Microtubule Architecture
Abstract
Introduction
Acknowledgments
References

22. Structure–Function Analysis of Yeast Tubulin
Abstract
Introduction
Acknowledgments
References

23. Using MTBindingSim as a Tool for Experimental Planning and Interpretation
Abstract
Introduction
Conclusion
References

24. Imaging Individual Spindle Microtubule Dynamics in Fission Yeast
Abstract
Introduction
Conclusion
Acknowledgments
References

Index
Volumes in Series