Barker C.J. (ed.) Inositol Phosphates and Lipids: Methods and Protocols

Humana, 2010. — 242 p.Inositol, in its native or lipid derived forms, serves as a master building block which, when phosphorylated, leads to the construction of more than 30 unique isomeric forms that are employed in vital but diverse regulatory roles in cells. In Inositol Phosphates and Lipids: Methods and Protocols, expert researchers introduce the basic methodological tools to measure inositol lipids and phosphates and also describe new approaches that have become available in the last 10 years, including RNA-silencing and the use of fluorescently labeled PH-domains to measure inositides in real-time in live cells, new sensitive methods to measure mass of both phosphates and lipids, as well as protocols involving inositol pyrophosphates. 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 notes sections, detailing tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Inositol Phosphates and Lipids: Methods and Protocols compiles many of the techniques that underscore phosphorylated inositol cell biology in one convenient guide.Preface
Contributors
The Role of Inositol and the Principles of Labelling, Extraction, and Analysis of Inositides in Mammalian Cells
HPLC Separation of Inositol Polyphosphates
Molecular Manipulation and Analysis of Inositol Phosphate and Pyrophosphate Levels in Mammalian Cells
A Femtomole-Sensitivity Mass Assay for Inositol Hexakisphosphate
Synthesis of InsP7 by the Inositol Hexakisphosphate Kinase 1 (IP6K1)
Protein Pyrophosphorylation by Diphosphoinositol Pentakisphosphate (InsP7)
Synthesis and Nonradioactive Micro-analysis of Diphosphoinositol Phosphates by HPLC with Postcolumn Complexometry
Diphosphosinositol Polyphosphates and Energy Metabolism: Assay for ATP/ADP Ratio
Isolation of Inositol 1,4,5-Trisphosphate Receptor-Associating Proteins and Selective Knockdown Using RNA Interfer
Phosphoinositide-Specific Phospholipase C b1 Signal Transduction in the Nucleus
Methods to Assess Changes in the Pattern of Nuclear Phosphoinositides
Measurement of Phosphoinositide 3-Kinase Products in Cultured Mammalian Cells by HPLC
Phosphoinositide Analysis by Liquid Chromatography–Mass Spectrometry
Imaging Phosphoinositide Dynamics in Living Cells
Index