The role of inositol and the principles of labelling, extraction, and analysis of inositides in mammalian cells / Christopher J. Barker and Per-Olof Berggren -- HPLC separation of inositol polyphosphates / Christopher J. Barker, Christopher Illies, and Per-Olof Berggren -- Molecular manipulation and analysis of inositol phosphate and pyrophosphate levels in mammalian cells / James C. Otto and John D. York -- A femtomole-sensitivity mass assay for inositol hexakisphosphate / Andrew J. Letcher, Michael J. Schell, and Robin F. Irvine -- Synthesis of InsP-- by the inositol hexakisphosphate kinase 1 (IP6K1) / Cristina Azevedo [and others] -- Protein pyrophosphorylation by diphosphoinositol pentakisphosphate (InsP-- ) / J. Kent Werner Jr., Traci Speed, and Rashna Bhandari -- Synthesis and nonradioactive micro-analysis of diphosphoinositol phosphates by HPLC with postcolumn complexometry / Hongying Lin, Karsten Lindner, and Georg W. Mayr -- Diphosphosinositol polyphosphates and energy metabolism : assay for ATP/ADP ratio / Andreas Nagel [and others] -- Isolation of inositol 1,4,5-triphosphate receptor-associating proteins and selective knockdown using RNA interference / Akihiro Mizutani [and others] -- Phosphoinositide-specific phospholipase C [beta]1 signal transduction in the nucleus / Roberta Fiume [and others] -- Methods to assess changes in the pattern of nuclear phosphoinositides / Nullin Divecha -- Measurement of phosphoinositide 3-kinase products in cultured mammalian cells by HPLC / Frank T. Cooke -- Phosphoinositide analysis by liquid chromatography-mass spectrometry / Trevor R. Pettitt -- Imaging phosphoinositide dynamics in living cells / Anne Wuttke, Olof Idevall-Hagren, and Anders Tengholm.
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SUMMARY OR ABSTRACT
Text of Note
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.