Microtubules, in vitro, Volume 115

Series Page

Contributors

Preface

Chapter 1. Site-Specific Fluorescent Labeling of Tubulin

Abstract

Introduction

Acknowledgments

References

Chapter 2. Measuring Microtubule Persistence Length Using a Microtubule Gliding Assay

Abstract

Introduction

Summary

Acknowledgments

References

Chapter 3. Structural Studies of the Doublecortin Family of MAPs

Abstract

Introduction

Acknowledgments

References

Chapter 4. Detection and Quantification of Microtubule Detachment from Centrosomes and Spindle Poles

Abstract

Introduction

Summary

Acknowledgments

References

Chapter 5. Regulation of Tubulin Expression by Micro-RNAs: Implications for Drug Resistance

Abstract

Introduction and Rationale

Summary

References

Chapter 6. Determining the Structure–Mechanics Relationships of Dense Microtubule Networks with Confocal Microscopy and Magnetic Tweezers-Based Microrheology

Abstract

Introduction

Summary

Acknowledgments

References

Chapter 7. Studying Mitochondria and Microtubule Localization and Dynamics in Standardized Cell Shapes

Abstract

Introduction

Conclusion

Acknowledgments

References

Chapter 8. Going Solo: Measuring the Motions of Microtubules with an In Vitro Assay for TIRF Microscopy

Abstract

Introduction

Concluding Remarks

Acknowledgments

References

Chapter 9. Analysis of Microtubules in Isolated Axoplasm from the Squid Giant Axon

Abstract

9.1 Preparation of Axoplasm

9.2 Analysis of Axoplasmic Microtubule Dynamics

9.3 Biochemistry of Axoplasmic Microtubules

9.4 Immunohistochemistry of Axoplasmic Microtubules

References

Chapter 10. Imaging GTP-Bound Tubulin: From Cellular to In Vitro Assembled Microtubules

Abstract

Introduction

Acknowledgments

References

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

Abstract

Introduction

References

Chapter 12. Heterotrimeric G Proteins and Microtubules

Abstract

Introduction

Concluding Comments

Acknowledgments

References

Chapter 13. Purification and Biophysical Analysis of Microtubule-Severing Enzymes In Vitro

Abstract

Introduction

Discussion and Summary

Acknowledgments

References

Chapter 14. Measurement of In Vitro Microtubule Polymerization by Turbidity and Fluorescence

Abstract

14.1 Background and Theory

14.2 Materials and Equipment

14.3 Methods

References

Chapter 15. Live-Cell Imaging of Microtubules and Microtubule-Associated Proteins in Arabidopsis thaliana

Abstract

Introduction

Concluding Comments

Acknowledgment

References

Chapter 16. Investigating Tubulin Posttranslational Modifications with Specific Antibodies

Abstract

Introduction

Concluding Remarks

Acknowledgments

References

Chapter 17. Purification and Assembly of Bacterial Tubulin BtubA/B and Constructs Bearing Eukaryotic Tubulin Sequences

Abstract

Introduction and Rationale

Conclusions

Acknowledgments

References

Chapter 18. Microtubule-Associated Proteins and Tubulin Interaction by Isothermal Titration Calorimetry

Abstract

Introduction

Conclusion

Acknowledgment

References

Chapter 19. Methods for Studying Microtubule Binding Site Interactions: Zampanolide as a Covalent Binding Agent

Abstract

Introduction

Summary

Acknowledgments

References

Chapter 20. Studying Kinetochore-Fiber Ultrastructure Using Correlative Light-Electron Microscopy

Abstract

Introduction

Acknowledgments

References

Chapter 21. Fluorescence-Based Assays for Microtubule Architecture

Abstract

Introduction

Acknowledgments

References

Chapter 22. Structure–Function Analysis of Yeast Tubulin

Abstract

Introduction

Acknowledgments

References

Chapter 23. Using MTBindingSim as a Tool for Experimental Planning and Interpretation

Abstract

Introduction

Conclusion

References

Chapter 24. Imaging Individual Spindle Microtubule Dynamics in Fission Yeast

Abstract

Introduction

Conclusion

Acknowledgments

References

Index

Volumes in Series

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 2e 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.

• Chapters reflect experimental procedures and new developments in the field of microtubule in vitro research
• Combines classical approaches and modern technologies
• Presents easy-to-use protocols and thorough background information, compiled by leaders in the field

Researchers and students in cell, molecular and developmental biology