Lipid Signaling Protocols

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 Biology™ 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.

Lipid Systems and Techniques.- Eicosanoids: Generation and Detection in Mammalian Cells.- Lipidomic Analysis of Phospholipids and Related Structures by Liquid Chromatography-Mass Spectrometry.- Measurement of Polyphosphoinositides in Cultured Mammalian Cells.- Inositol Lipid-Dependent Functions in : Analysis of Phosphatidylinositol Phosphates.- Methods for the Determination of the Mass of Nuclear PtdIns4, PtdIns5, and PtdIns(4,5).- Lipid Quantification and Structure Determination of Nuclear Envelope Precursor Membranes in the Sea Urchin.- Solution and Solid-State NMR of Lipids.- Spatial Localization of PtdIns in Phase-Separated Giant Unilamellar Vesicles with a Fluorescent PLC-delta 1 PH Domain.- Studying Cell-to-Cell Interactions: An Easy Method of Tethering Ligands on Artificial Membranes.- Ceramide-Induced Transbilayer (Flip-Flop) Lipid Movement in Membranes.- Effects of Sterols on the Development and Aging of.- Proteomic Analysis of the Sterol-Mediated Signaling Pathway in.- Lipid Metabolism and Recognition.- Studying the Subcellular Localization and DNA-Binding Activity of FoxO Transcription Factors, Downstream Effectors of PI3K/Akt.- Measurement of PTEN Activity by Imaging Phosphorylated Akt.- Analysis of Phosphatidylinositol 3,4,5 Trisphosphate 5-Phosphatase Activity by and Assays.- Measuring Phospholipase D Activity in Insulin-Secreting Pancreatic #x03B2;-Cells and Insulin-Responsive Muscle Cells and Adipocytes.- Protein Kinase C as an Effector of Lipid-Derived Second Messengers.- Detection of Myotubularin Phosphatases Activity on Phosphoinositides and.- Quantification of Multiple Phosphatidylinositol 4-Kinase Isozyme Activities in Cell Extracts.- Phospholipid-Interacting Proteins by Solution-State NMR Spectroscopy.- Revealing Signaling in Single Cells by Single- and Two-Photon Fluorescence Lifetime Imaging Microscopy.- Phosphoinositide (PI) 3-Kinase Assays.- Measurement of Phosphatidylinositol and Phosphatidylcholine Binding and Transfer Activity of the Lipid Transport Protein PITP.- Reconstitution of Activation of PLC#x03B5; by Ras and Rho GTPases.- Assaying Endogenous Phosphatidylinositol-4-Phosphate 5-Kinase (PIP5K) Activities.- Preparation of Membrane Rafts.