Lipid Signaling Protocols

In this second edition methods are described to measure the synthesis of lipids such as the phosphoinositides, ceramides and sphingomyelin, as well as techniques to molecularly characterize the various kinases and phosphatases that regulate the intracellular metabolism of these lipids. Lipid Signaling Protocols, Second Edition guides readers through detailed experimental protocols, which are complimented by review chapters that highlight the technical considerations, challenges and potential pitfalls associated with using these laboratory-based approaches. Written for the Methods in Molecular Biology series, 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.

Authoritative and practical, Lipid Signaling Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Method for Assaying the Lipid Kinase Phosphatidylinositol-5-phosphate 4-kinase in Quantitative High-Throughput Screening (qHTS) Bioluminescent Format.- Assaying Ceramide Synthase Activity In Vitro and in Living Cells using Liquid Chromatography-Mass Spectrometry.- Fluorescent Assays for Ceramide Synthase Activity.- Identification of the Interactome of a Palmitoylated Membrane Protein, Phosphatidylinositol 4-kinase type II Alpha.- Measurement of Long Chain Fatty Acyl-CoA synthetase Activity.- Qualitative and Quantitative in vitro Analysis of Phosphatidylinositol Phosphatase Substrate Specificity.- Luciferase Reporter Assays to Assess Liver X Receptor Transcriptional Activity.- Metabolically Biotinylated Reporters for Electron Microscopic Imaging of Cytoplasmic Membrane Microdomains.- Fluorescence Recovery after Photobleaching Analysis of the Diffusional Mobility of Plasma Membrane Proteins: HER3 Mobility in Breast Cancer Cell Membranes.- Isolation and Analysis of Detergent-Resistant Membrane Fractions.- Detection of Isolated Mitochondria-Associated ER Membranes using the Sigma-1 Receptor.- Using Surface Plasmon Resonance to Quantitatively Assess Lipid-Protein Interactions.- Analyzing Protein-phosphoinositide Interactions with Liposome Flotation Assays.- High-Throughput Flurometric Assay for Membrane-Protein Interaction.- Guidelines for the use of Protein Domains in Acidic Phospholipid Imaging.- Analysis of Sphingolipid Synthesis and Transport by Metabolic Labeling of Cultured Cells with [ 3 H]serine.- Determination and Characterization of Tetraspanin-Associated Phosphoinositide-4 Kinases in Primary and Neoplastic Liver Cells.- Analysis of the Phosphoinositide Composition of Subcellular Membrane Fractions.- Single-molecule Imaging of Signal Transduction via GPI-anchored Receptors.- Measuring Phosphatidylinositol Generation on Biological Membranes.- Assay for CDP-diacylglycerol Generation by CDS in Membrane Fractions.