As scientist begin to understand the complexity of lipid signaling and its roles in plant biology, there is an increasing interest in their analysis. Due to the low abundancy and transient nature of some of these hydrophobic compounds, this is not always easy. In Plant Lipid Signaling Protocols, expert researchers in the field detail experimental approaches by which plant signaling lipids can be studied. These methods and techniques include analysis of plant signaling lipids, including detailed protocols to detect various relevant compounds by targeted or non-targeted approaches; to assay relevant enzyme activities in biological material or using recombinant enzymes; to test for specific binding of signaling lipids to protein partners; or to visualize signaling lipids or lipid-derived signals in living plant 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Plant Lipid Signaling Protocols aids plant researchers in the continuing to study the roles of lipid signals.

The previous edition of Transmembrane Signaling Protocols was published in 1998. Since then the human genome has been completely sequenced and new methods have been developed for the use of microarrays and proteomics to analyze global changes in gene expression and protein profiles. These advances have increased our ability to understand transmembrane signaling processes in much greater detail. They have also simultaneously enhanced our ability to determine the role of a large number of newly identified molecules in signaling events. In addition, novel video microscopy methods have been developed to image transmembrane signaling events in live cells in real time. In view of these major advances, it is time to update the previous edition. Because of the success of that volume, we have chosen to keep the essential character of the book intact. Introductory chapters from experts have been included to provide overall perspective and an overview of recent advances in signal transduction pathways. The individual chapters now include comp- hensive detailed methods, studies in genetically tractable systems, fluorescence microscopy in live single cells, ex vivo analysis of primary cells from tra- genic mice, as well as genomic and proteomic approaches to the analysis of transmembrane signaling events. We would like to express our deep gratitude to the coauthors of this publi- tion. We hope that Transmembrane Signaling Protocols, Second Edition will serve as a valuable resource for future progress in the study of signal transd- tion pathways.

This collection of practical, cutting-edge techniques for the study of cell signaling provides detailed, step-by-step instructions, helpful notes, and troubleshooting tips that make even the most powerful of the newest techniques readily reproducible. The protocols presented include the use of peptide libraries to study transmembrane signaling; the use of single-cell assays to analyze signal transduction pathways; the reconstitution of signaling complexes; methods for analyzing protein-protein interactions, and more. Introductory reviews explain the basic theory and enable researchers new to the area to rapidly gain understanding, as well as command of the practical knowledge and expertise afforded by the protocols. Transmembrane Signaling Protocols makes available to all researchers the many state-of-the-art techniques that have recently led to landmark discoveries in transmembrane signaling.

In the first edition of Calcium Signaling Protocols I began by writing “The regula- 2+ tion of intracellular Ca is a common theme presented in many papers over the last 20 2+ or so years and the description of the Ca -sensitive indicator dye fura-2 in 1985 resulted in a massive increase in these types of studies. ” This statement is as true in 2005 as it was in 1999, but 20 or so years is now 30 years! There has been some reorganization of the volume such that there are now 22 ch- ters including five new ones, all written by experts in their field. These new chapters 2+ include use of the FlexStation and electrophysiological measurement of Ca channel activity. The book is broken into six parts. Part I is a general coverage of basic theory and the simplest use of fluorescent indicators. Part II covers specialist measurement 2+ systems and Part III covers measurement of Ca channel activity. Assessment of 2+ release of stored Ca is covered in some detail in Part IV, with Parts V and VI cover- 2+ ing specialist measurement techniques and Ca -sensitive targets. Putting a book like this together, even as a second edition, takes time and I am, again, indebted to the individual authors for their help and patience. I am also very grateful to Professor John M. Walker, the series editor, for his continued help and advice over the course of this project.

This volume explores analytical methods to study complex lipid mixtures from plants and algae. The chapters in this book are organized into five parts and cover topics such as basic methods of lipid isolation and analysis; mass spectrometry and NMR analysis; lipid isolation and analysis from plant tissues, cell compartments and organelles; lipid signaling, lipid-protein interactions, and imaging; and lipid databases. 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. Cutting-edge and comprehensive, Plant Lipids: Methods and Protocols is a valuable guide for experienced researchers and undergraduate, graduate, and Ph.D. students. This book is also an excellent resource for novice scientists with little to no experience in lipid experiments who are interested in approaching this field experimentally.

Lipid Signaling Protocols assembles in a single volume the various tools and methodologies needed by the interested investigator to unravel lipid dependent signaling and cell function. Divided into two convenient sections, the volume begins by summarizing the physical properties of hydrophobic metabolites as well as the physical methodologies used for their analysis, which leads to the second section and its selection of biological methods, focused around the most relevant lipids, their corresponding metabolizing enzymes and the recognition proteins. Following the highly successful Methods in Molecular BiologyTM series format, the chapters provide readily reproducible laboratory protocols, lists of necessary materials and reagents, and the tips on troubleshooting and avoiding known pitfalls. Contributed to by top researchers in the field, Lipid Signaling Protocols is an essential resource for both experienced and novice researchers who desire a better understanding of the application of physical methodologies in the context of lipid signaling and lipid metabolism in cell biology.