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.