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.

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.

In Phospholipid Signaling Protocols, state-of-the-art techniques for monitoring the major lipid and phospholipid-derived second messenger pathways to identify and quantify pathway activation are detailed by experts intimately experienced in their use. The assays described cover all the major phospholipases (C, D, A2), as well as sphingomyelinase and its associated metabolites. Additional protocols are provided for the assay of phosphoinositide 3-, 4-, and 5-kinase and sphingosine kinase activity, and for the quantification, separation, and rigorous identification of phospholipids, diacylglycerol, and sphingolipids, as well as their metabolites, including phosphoinositols, choline metabolites, and fatty acid metabolites. In addition, there is extensive information on the extraction, size separation, detection, and quantification of cellular signaling proteins and corresponding mRNA, as well as a description of their localization by immunohistochemistry and immunocytochemistry. Phospholipid Signaling Protocols offers a wide-ranging collection of cutting-edge techniques for the study of signal transduction through phospholipid intermediates and their metabolites. The book is an indispensable reference for both the newcomer and the experienced researcher seeking to expand knowledge of these critical pathways, and strongly complements its companion volumes-R.A.J. Challiss' Receptor Signal Transduction Protocols, D. Bar-Sagi's, Transmembrane Signaling Protocols, and D. A. Kendall and S.J. Hill's Signal Transduction Protocols-in building a unique library of tried-and-tested protocols relating to the ever expanding signal transduction field.

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.

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.

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.