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This second edition of Receptor Signal Transduction Protocols not only has a new editor, but also a greater focus on G-protein-coupled receptors, their properties per se, and their coupling to immediate downstream binding partners—principally, although not exclusively, the heterotrimeric G-proteins. The new edition combines updates of key chapters from the first edition, as well as a large number of new contributions covering key methodologies that have emerged, or been extended to receptor/G-protein research, in the past 5–6 years. In common with many fields, the range of methods used to assess the first steps in signal transduction are continually expanding and methods that might have been considered too specialized five years ago are now sufficiently routine to be included here. Unlike many research areas, where off-the-shelf kits have made research basically foolproof, signal transduction research still requires considerable expertise, and the methods included here are provided by internationally recognized experts in their fields who have many years of experience using the methods they describe. This not only allows each chapter to impart a clear description of the method, but also to furnish invaluable troubleshooting advice for when things do not go entirely according to plan. Once again we would like to thank the Series Editor, John Walker, for the invitation to compile this second edition, and to express our gratitude to all of the authors who have enthusiastically agreed to provide the uniformly excellent contributions.
This fully updated volume reflects the spectacular advances in our knowledge of signal transduction pathways with a selection of ‘classic’ as well as newly developed approaches. These detailed approaches expand into the fields of molecular biology, biochemistry, physiology, cell biology, genetics, and genomics. 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. Practical and up-to-date, Plant Signal Transduction: Methods and Protocols, Second Edition serves as an ideal guide for researchers exploring the vast array of signals produced by plants to ensure their survival.
During recent years, research has greatly expanded our understanding of the sophisticated molecular network of responses which enable plants to develop, survive and propagate under a wide range of conditions. In Plant Signal Transduction: Methods and Protocols, an international panel of experts provide well-established methods vital to analyzing plant signal transduction on the molecular level. Featuring experimental procedures on several of the most popular model organisms, the volume focuses on in planta analyses and the proteins involved in signal transduction in order to aid with the establishment of laboratory techniques or the modification of the protocols for other plants. As part of the highly successful Methods in Molecular BiologyTM series, the chapters include brief introductions to the subject, lists of necessary materials, readily reproducible laboratory protocols, and tips on trouble-shooting and avoiding known pitfalls. Comprehensive and cutting-edge, Plant Signal Transduction: Methods and Protocols will benefit plant scientists wishing to improve their experimental approaches and delve further into this exciting and important field of study
"This textbook provides a comprehensive view of signal transduction, covering both the fundamental mechanisms involved and their roles in key biological processes. It first lays out the basic principles of signal transduction, explaining how different receptors receive information and transmit it via signaling proteins, ions, and second messengers. It then surveys the major signaling pathways that operate in cells, before examining in detail how these function in processes such as cell growth and division, cell movement, metabolism, development, reproduction, the nervous system, and immune function"--
In this second edition of a widely appreciated work, Receptor Signal Transduction Protocols, a panel of internationally recognized investigators presents their best methods for studying G-protein-coupled receptors (GPCRs) and events immediately downstream of their activation. This new edition combines updates of key chapters from the first edition with a large number of new contributions on the many successful methodologies that have emerged more recently. The methods are focused primarily on events at the receptor level, including ligand binding, on the genetic manipulation of receptors, the generation of model cell lines in which to study them, and the interaction and activation of G-proteins. Additional methods concentrate on receptor expression and localization, receptor internalization and post-translational modification, GPCR-protein interactions, and the use of knock-out and knock-in strategies for determining the physiological roles of receptors. The laboratory protocols follow the successful Methods in Molecular Biology"!series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and highly practical, Receptor Signal Transduction Protocols, Second Edition offers both novice and experienced investigators powerful cutting-edge techniques that provide an array of approaches and specific methods that will aid in the understanding of GPCR structure and function
As our understanding of the biological sciences expands, the bou- aries between traditional disciplines tend to blur at the edges. Physio- gists and pharmacologists, for instance, now need to embrace techniques that until recently were the strict preserves of biochemists and mole- lar biologists. However, the acquisition of new technologies can be a time-consuming and frustrating business, and unless an expert is on hand to give instruction, precious hours can be spent poring over half-described Methods sections with no guarantee of eventual success. The aim of Signal Transduction Protocols has been to get experts with "hands-on" experience in particular techniques to give detailed accounts of experimental protocols in a recipe-type format, which we hope will circumvent the problems of ambiguity often encountered when reading the literature. The techniques described in Signal Transduction Protocols are those that we think will be most useful in addressing questions in the area of receptor-mediated cell signaling, with particular regard to those receptors that are part of the G-protein-linked superfamily. To keep it to a manageable size, we have omitted any reference to electrophysi- ogy and have instead concentrated on more biochemical approaches.
As our understanding of G protein-coupled receptor (GPCR) signal transduction continues to grow, we cannot help but be struck by the emerging complexity and the ability of this receptor superfamily to continually surprise us as new facets are discovered. In this third edition of Receptor Signal Transduction Protocols, the expert contributors have taken into account the constant evolution of the GPCR field and dealt with methods that allow researchers to address many of the important rising issues. Serving as a companion to the previous editions, this volume contains thorough revisions of some core chapters, the complete rewriting of others to encompass new technological developments since the publication of the prior editions, and the commissioning of brand new, cutting-edge chapters to expand on previous coverage. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to the 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 up-to-date, Receptor Signal Transduction Protocols, Third Edition serves the scientific community with a broad collection of the current, practical methodologies in this highly promising field.
In 1995, Signal Transduction Protocols, edited by David A. Kendall and Stephen J. Hill, was published in the Methods in Molecular Biology series. This second edition represents an update to that previous work with an emp- sis on new methodologies that have developed in the last few years. The goal, then and now, is to provide procedures written by experts with first-hand ex- rience in a detail that goes far beyond what is generally encountered in the “methods” section of most journals and thus actually permits a particular p- cedure to be replicated. In addition, we have had as a secondary goal the id- tification of protocols for the assay of general classes of signal transduction components that, ideally, can be adapted to the assay of any member of that class. The ability to do this has resulted in large part from the use of affini- based assays, the ease with which specific proteins can be specifically tagged, and an explosion in the availability of highly specific antibodies from comm- cial sources, especially antibodies raised against signaling proteins of human origin. The number of available approaches is, fortunately for those working in signaling research, far too great to fit within the confines of this volume, so hard choices as to what to include had to be made.
The rapid expansion of the TRP field has generated a large amount of excellent original work across many different research fields. However, investigators are not necessarily familiar with the pros and cons of the variety of methods used to study TRP channels. Because of functional and genetic diversity, as well as the different physiological roles
In Natural Killer Cell Protocols: Cellular and Molecular Methods, Kerry S. Campbell and Marco Colonna have assembled a comprehensive collection of readily reproducible methods designed to study natural killer (NK) cells from the broadest variety of viewpoints. These include not only classic techniques, but also new approaches to standard methods, newly evolved techniques that have become valuable for specific applications, and unique models for manipulating and studying NK cells. Among the advanced methods covered are those for in vitro transendothelial migration, in vivo detection of cells migrating into tumors, immunofluorescence staining of intracellular cytokines, and in vitro NK cell development. Valuable techniques for specific applications include vaccinia virus protein expression, soluble KIR-Fc fusions for HLA class I binding assays, calcium mobilization in cell conjugates, and identification of heterodimeric receptor complexes using cDNA library expression cloning. No less important are accounts of such classic methods as hybrid resistance, ADCC, viral defense, target cell cytotoxicity assays, cloning and culturing, tumor immunotherapy, and generation of HLA class I transfected target cells. Natural Killer Cell Protocols: Cellular and Molecular Methods offers immunologists, cancer researchers, virologists, and cell biologists today's most comprehensive collection of both established and cutting-edge techniques, methods that will contribute significantly to advancing our understanding of this fascinating and critically important class of cells.