Lipid Signaling Protocols (2^nd Ed., Softcover reprint of the original 2nd ed. 2016)
Methods in Molecular Biology Series, Vol. 1376

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.

1. Method for Assaying the Lipid Kinase Phosphatidylinositol-5-phosphate 4-kinase α in Quantitative High-Throughput Screening (qHTS) Bioluminescent Format

Mindy I. Davis, Atsuo T. Sasaki, and Anton Simeonov

2. Assaying Ceramide Synthase Activity In Vitro and in Living Cells using Liquid Chromatography-Mass Spectrometry

Xin Ying Lim, Russell Pickford, and Anthony S Don

3. Fluorescent Assays for Ceramide Synthase Activity

Timothy A Couttas and Anthony S Don

4. Identification of the Interactome of a Palmitoylated Membrane Protein, Phosphatidylinositol 4-kinase type II Alpha

Avanti Gokhale, Pearl V. Ryder, Stephanie A. Zlatic, and Victor Faundez

5. Measurement of Long Chain Fatty Acyl-CoA synthetase Activity

Joachim Füllekrug and Margarete Poppelreuther

6. Qualitative and Quantitative in vitro Analysis of Phosphatidylinositol Phosphatase Substrate Specificity

Laura Ren Huey Ip and Christina Anja Gewinner

7. Luciferase Reporter Assays to Assess Liver X Receptor Transcriptional Activity

Matthew C. Gage, Benoit Pourcet, and Inés Pineda-Torra

8. Metabolically Biotinylated Reporters for Electron Microscopic Imaging of Cytoplasmic Membrane Microdomains

Kimberly J. Krager and John G. Koland

9. Fluorescence Recovery after Photobleaching Analysis of the Diffusional Mobility of Plasma Membrane Proteins: HER3 Mobility in Breast Cancer Cell

Mitul Sarkar and John G. Koland

10. Isolation and Analysis of Detergent-Resistant Membrane Fractions

Massimo Aureli, Sara Grassi, Sandro Sonnino, and Alessandro Prinetti

11. Detection of Isolated Mitochondria-Associated ER Membranes using the Sigma-1 Receptor

Abasha Lewis and Tsung-Ping Su

12. Using Surface Plasmon Resonance to Quantitatively Assess Lipid-Protein Interactions

Kathryn Del Vecchio and Robert V. Stahelin

13. Analyzing Protein-phosphoinositide Interactions with Liposome Flotation Assays

Ricarda A. Busse, Andreea Scacioc, Amanda M. Schalk, Roswitha Krick, Michael Thumm, and Karin Kühnel

14. High-Throughput Flurometric Assay for Membrane-Protein Interaction

Wonhwa Cho, Hyunjin Kim, and Yusi Hu

15. Guidelines for the use of Protein Domains in Acidic Phospholipid Imaging

 Matthieu Pierre Platre and Yvon Jaillais

16. Analysis of Sphingolipid Synthesis and Transport by Metabolic Labeling of Cultured Cells with [³H]serine

Neale D. Ridgway

17. Determination and Characterization of Tetraspanin-Associated Phosphoinositide-4 Kinases in Primary and Neoplastic Liver Cells

Krista Rombouts and Vinicio Carloni

18. Analysis of the Phosphoinositide Composition of Subcellular Membrane Fractions

Deborah A. Sarkes  and Lucia E. Rameh

19. Single-molecule

Kenichi G. N. Suzuki

20. Measuring Phosphatidylinositol Generation on Biological Membranes

Mark G. Waugh

21. Assay for CDP-diacylglycerol Generation by CDS in Membrane Fractions

Mark G. Waugh

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts