Microtubules, in vitro (2^nd Ed.)
Methods in Cell Biology 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 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.

1. Isolation of Antarctic fish tubulin and MAPs
2. Making, isolating and characterizing tubulin alpha antibodies
3. Techniques for labeling tubulin and drugs
4. Single-particle 3D reconstruction methods for decorated and undecorated microtubules
5. A new microtubule gliding assay and analysis of microtubule persistence length
6. Characterization of gamma tubulin
7. Biochemical and Structural Studies of the doublecortin family of MAPs
8. Detection and quantification of microtubule detachment from centrosomes and spindle poles
9. Regulation of tubulin expression by micro RNAs: Implications for Drug Resistance
10. A portable magnetic tweezers device to visualize three-dimensional deformation of reconstituted microtubule networks
11. Mitochondrial/microtubule interactions – mmbp1
12. Going Solo: Measuring the motions of microtubules with an in vitro assay for TIRF microscopy
13. Analysis of Microtubules and Motors in Isolated Axoplasm from the Squid Giant Axon
14. Analysis of drug binding to microtubules and steady state microtubule growth/treadmilling by use of a sedimentation assay in vitro
15. Specific recombinant antibody technology for the study of microtubule dynamics and regulatory proteins
16. Tubulin-specific Chaperones: Components of a Molecular Machine that Assembles the aB Heterodimer
17. G-protein tubulin interactions
18. Katanin, a microtubule severing enzyme: expression, purification, and biophysical characterization
19. Measuring tubulin polymerization in vitro – comparison of methods and possible artifacts
20. Imaging microtubules in living plant cells
21. How to use which antibody known in the field, what to expect from this antibody and how semi-quantitative data can be reliably obtained with antibody staining
22. Preparation of bacterial tubulin BtubA/B and constructs bearing eukaryotic tubulin sequences
23. Microtubule-associated proteins and tubulin interaction by isothermal titration calorimetry
24. TBC
25. Characterization of the mechanism of action of the covalent-binding microtubule-stabilizing agent zampanolide
26. Studying kinetochore-fibre ultrastructure using correlative light-electron microscopy
27. Fluorescence-based Assays for Microtubule Structure
28. Yeast Tubulin-structure-function analysis
29. Microtubule Binding SIm to simulate binding of proteins to polymers under different binding models

University of California, Santa Barbara, USA