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.

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.

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.

Since the publication of the first edition of Signal Transduction: A Practical Approach in 1992 there has been a great deal of new information about the processes of signal transduction and consequently many new methods have been developed. This new edition has therefore been updated and extended to include the major new methods now available. The first part of the book is mainly concerned with G protein-coupled receptors and covers structural studies of conformational changes and binding sites, phosphorylation and desensitisation, identification, receptor fusion proteins, and reporter gene systems. The second part includes methods for studying components of the other major families of signal transduction: adenylyl cylase and cAMP, phosphorylated inositol lipids, phosphinositide 3-kinases, phosphlipase D and phosphatidylcholine, sphingosine kinase, and inositol 1,4,5-triphosphate. Also included are chapters on baculoviral expression systems and the quantitative assay of mitogen activated protein kinases in intact cells and tissues. As with the previous edition Signal Transduction 2e covers a wide range of techniques and will be useful to both experienced researchers and newcomers.

Signalling within and between cells is an essential part of many biological processes, from the development of the body to the activity of our immune system. Cell Signalling, Third Edition, presents a carefully structured introduction to this intricate subject, introducing those conserved features that underlie many different extra- and intracellular signalling systems. Starting with an overview of cell signalling and highlighting its importance in many biological systems, the book goes on to explore the key components of extracellular and intracellular signalling mechanisms before examining how these components come together to create signalling pathways, which are so crucial to the survival of many living organisms. The text is enhanced by two-color artwork and 3-D protein models. A Companion Website provides resources for students and instructors.

PCR in Bioanalysis offers powerful PCR-based protocols and assays in actual use or potential use in clinical medicine and commercial biology. The main focus of the book is on the commercial applications of PCR, as opposed to basic research uses. Topics covered include the measurement of hormone levels using PCR, transcription factor isolation, detection of viruses using PCR, detection of tumor contamination of stem cells, evaluation of grafts for tumor cells, and more.

Intracellular checkpoint controls constitute a network of signal transd- tion pathways that protect cells from external stresses and internal errors. Ext- nal stresses can be generated by the continuous assault of DNA-damaging agents, such as environmental mutagens, ultraviolet (UV) light, ionizing radiation, or the reactive oxygen species that can arise during normal cellular metabolism. In response to any of these assaults on the integrity of the genome, the activation of the network of checkpoint control pathways can lead to diverse cellular responses, such as cell cycle arrest, DNA repair, or elimination of the cell by cell death (apoptosis) if the damage cannot be repaired. Moreover, internal errors can occur during the highly orchestrated replication of the cellular genome and its distribution into daughter cells. Here, the temporal order of these cell cycle events must be strictly enforced—for example, to ensure that DNA replication is c- plete and occurs only once before cell division, or to monitor mitotic spindle assembly, and to prevent exit from mitosis until chromosome segregation has been completed. Thus, well functioning checkpoint mechanisms are central to the maintenance of genomic integrity and the basic viability of cells and, the- fore, are essential for proper development and survival. The importance of proper functioning of checkpoints becomes plainly obvious under conditions in which this control network malfunctions and fails. Depending on the severity and timing, failure of this machinery can lead to embryonic lethality, genetic diseases, and cancer.

Gazing into crystal balls is beyond the expertise of most scientists. Yet, as we look further into the 21st century, one does not have to be Nostradamus to predict that the current genomics and proteomics "revolution" will have an immense impact on medical bacteriology. This impact is already being re- ized in many academic departments, and although encroachment on routine diagnostic bacteriology, particularly in the hospital setting, is likely to occur at a slower pace, it remains nonetheless inevitable. Therefore, it is important that no one working in bacteriology should find themselves distanced from these fundamental developments. The involvement of all clinical bacteriologists is essential if the significant achievements of genome sequencing and analysis are to be turned into tangible advances, with resulting benefits for patient care and m- agement. It is our hope that Genomics, Proteomics, and Clinical Bacteriology: Methods and Reviews will play a part in bringing such a development to fruition. The advances in genomics and proteomics have already given us frequent opportunities to reassess our knowledge and understanding of established b- terial adversaries, and have provided us with the means to identify new foes. The new knowledge gained is enabling us to reconsider, for example, our c- cepts of bacterial pathogenicity, phylogeny and novel targets for antibacterial chemotherapy. These topics, and others, are considered in Genomics, Proteomics, and Clinical Bacteriology: Methods and Reviews.

In Confocal Microscopy Methods and Protocols, Stephen Paddock and a highly skilled panel of experts lead the researcher using confocal techniques from the bench top, through the imaging process, to the journal page. They concisely describe all the key stages of confocal imaging-from tissue sampling methods, through the staining process, to the manipulation, presentation, and publication of the realized image. Written in a user-friendly, nontechnical style, the methods specifically cover most of the commonly used model organisms: worms, sea urchins, flies, plants, yeast, frogs, and zebrafish. Centered in the many biological applications of the confocal microscope, the book makes possible the successful imaging of both fixed and living specimens using primarily the laser scanning confocal microscope. The powerful hands-on methods collected in Confocal Microscopy Methods and Protocols will help even the novice to produce first-class cover-quality confocal images.

A comprehensive collection of readily reproducible methods for studying receptors in silico, in vitro, and in vivo. These cutting-edge techniques cover mining from curated databases, identifying novel receptors by high throughput screening, molecular methods to identify mRNA encoding receptors, radioligand binding assays and their analysis, quantitative autoradiography, and imaging receptors by positron emission tomography (PET). Highlights include phenotypic characterization of receptors in knockout mice, imaging receptors using green fluorescent protein and fluorescent resonance energy transfer, and quantitative analysis of receptor mRNA by TaqMan PCR. These book equips the researcher with techniques for exploring the unprecedented number of new receptor systems now emerging and the so-called "orphan" receptors whose activating ligand has not been identified.