Microtubules are essential components of the cytoskeleton, and play critical roles in a variety of cell processes, including cell shaping, intracellular tracking, cell division, and cell migration. Microtubule Protocols presents a comprehensive collection of essential and up-to-date methods for studying both the biology of microtubules and the mechanisms of action of microtubule-interacting drugs. The straightforward presentation of readily reproducible protocols is a hallmark of the Methods in Molecular MedicineTM series, and is evident in this volume. Methods presented range from the purification and characterization of microtubule proteins, analysis of post-translational modifications of tubulin, and determination of microtubule structure, to the visualization of microtubule and spindle behavior, measurement of microtubule dynamics, and examination of microtubule-mediated cellular processes. Both basic scientists and clinical researchers will benefit from this collection of state-of-the-art protocols for microtubule research.

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 BiologyTM 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.

This volume provides state-of-the-art methodology to functionalize microtubules and construct microtubule-based materials. Chapters consists of six sections detailing modification of microtubule structures, observation and control of microtubule movement, material applications of microtubules in vitro, development of microtubule-binding molecules, comprehensive approaches to analyze properties of microtubules, and functionalization of microtubules in living cells. 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. Authoritative and cutting-edge, Microtubules: Methods and Protocols aims to provide useful information for construction of microtubule-based materials and help to create new ideas for the next-generation of materials and applications through detailed protocols.

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 volume (complemented by the forthcoming companion volume by Cassimeris and Tran) brings together current research and protocols in the field of microtoubules 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 both 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

This volume details comprehensive state-of-the-art methods on actin microfilaments and microtubules and how they work to achieve different cellular functions in different cellular contexts. Chapters guide readers through protein purification, in vitro reconstitution of several cytoskeleton properties, analyses of microtubule- and actin-based structures, functional dissection of post-translational modifications, and roles in several biological processes. 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. Authoritative and cutting-edge, Cytoskeleton Dynamics: Methods and Protocols aims to provide a wide range of experimental approaches and be an invaluable resource for present and future generations of cytoskeleton researchers. The chapter “Visualization and Functional Analysis of Spindle Actin and Chromosome Segregation in Mammalian Oocytes” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

The decision, in 1975, to write alone a monograph on micro tubules was not without risks. While I was familiar from its start in Brussels in 1934 with the work on col chicine and other mitotic poisons, the literature on microtubules was, 8 years ago, already increasing at an impressive rate. However, this monograph, which, contrary to other works on microtubules, tried to cover the whole field of research, from the fundamentals of the tubulin molecule and the possible role of these organelles in some aspects of human pathology, to some medical applications of microtubule poisons, has been accepted as a useful tool for workers in these fields. Since 1976, (date of the last references mentioned in the monograph) until the middle of 1983, papers on microtubule research have literally been pouring in, at the rate of several hundred a year. This may justify a second edition, although the considerable difficulties in keeping the size of the book within the same limits while not forgetting to mention some important work, could not be overlooked. The need for an entirely revised and rewritten edition prompted this new venture and was possible with the help of the considerable amount of reprints kindly sent to me day after day over the years. This work would have been unthinkable if the author had not maintained the same enthusiasm for microtubule research, which has been disclosing new facts every day.

Microtubules: in vivo includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton. The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation. - Includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton - The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation

In the last 10 years researchers have firmly established key roles for R- related GTPases in almost every aspect of cell biology. In the 1980s the pro- oncogene Ras itself was the focus of interest, though in the 1990s this shifted to the increasing variety of Ras-related proteins. In this new decade much yet needs to be done to establish the role for all the small GTPases now uncovered by the human genome project. In particular, these GTPases need to be und- stood in the appropriate biochemical and cellular contexts. In the process of trying to uncover the role of these versatile proteins, a variety of novel te- niques and methodologies has been developed. These now enable investi- tors to move easily within a diversity of fields ranging from structural studies to real-time in vivo analysis of a GTPase. In recognition of the need for access to key background methodologies, GTPase Protocols: The Ras Superfamily is devoted to techniques that are pr- ently widely used and that will continue to be the standard for researchers worldwide. Each chapter is aimed at supplying detailed methodologies to allow reproduction in any laboratory, while also providing the general pr- ciples on which the methods are based. Some of the techniques grouped in the first section apply broadly to small GTPases, whereas others in Part II are more applicable within each GTPase subfamily.

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. - 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

This volume provides state-of-the-art methodology to functionalize microtubules and construct microtubule-based materials. Chapters consists of six sections detailing modification of microtubule structures, observation and control of microtubule movement, material applications of microtubules in vitro, development of microtubule-binding molecules, comprehensive approaches to analyze properties of microtubules, and functionalization of microtubules in living cells. 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. Authoritative and cutting-edge, Microtubules: Methods and Protocols aims to provide useful information for construction of microtubule-based materials and help to create new ideas for the next-generation of materials and applications through detailed protocols.