Inositol phosphates and lipids : methods and protocols
著者
書誌事項
Inositol phosphates and lipids : methods and protocols
(Methods in molecular biology / John M. Walker, series editor, 645)(Springer protocols)
Humana, 2010
- : hbk
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注記
Includes bibliographical references and index
内容説明・目次
内容説明
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 (TM) 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.
目次
Table of Contents
Preface
Contributors
1. The Role of Inositol and the Principals of Labelling, Extraction, and Analysis of Inositides in Mammalian Cells
Christopher J. Barker and Per-Olof Berggren
2. HPLC Separation of Inositol Polyphosphates
Christopher J. Barker, Christopher Illies, and Per-Olof Berggren
3. Molecular Manipulation and Analysis of Inositol Phosphate and Pyrophosphate Levels in Mammalian Cells
James C. Otto and John D. York
4. A Femtomole-Sensitivity Mass Assay for Inositol Hexakisphosphate
Andrew J. Letcher, Michael J. Schell, and Robin F. Irvine
5. Synthesis of InsP7 by the Inositol Hexakisphosphate Kinase 1 (IP6K1)
Cristina Azevedo, Adam Burton, Matthew Bennett, Sara Maria Nancy Onnebo, and Adolfo Saiardi
6. Protein Pyrophosphorylation by Diphosphoinositol Pentakisphosphate (InsP7)
J. Kent Werner, Jr., Traci Speed, and Rashna Bhandari
7. Synthesis and Non-Radioactive Micro-Analysis of Diphosphoinositol Phosphates by HPLC with Post-Column Complexometry
Hongying Lin, Karsten Lindner, and Georg W. Mayr
8. Diphosphosinositol Polyphosphates and Energy Metabolism: Assay for ATP/ADP Ratio
Andreas Nagel, Christopher J. Barker, Per-Olof Berggren, and Christopher Illies
9. Isolation of Inositol 1,4,5-Trisphosphate Receptor-Associating Proteins and Selective Knockdown Using RNA Interference
Akihiro Mizutani, Kastuhiro Kawaai, Chihiro Hisatsune, Hideaki Ando, Takayuki Michikawa, and Katsuhiko Mikoshiba
10. Phosphoinositide-Specific Phospholipase C b1 Signal Transduction in the Nucleus
Roberta Fiume, Gabriella Teti, Irene Faenza, and Lucio Cocco
11. Methods to Assess Changes in the Pattern of Nuclear Phosphoinositides
Nullin Divecha
12. Measurement of Phosphoinositide 3-Kinase Products in Cultured Mammalian Cells by HPLC
Frank T. Cooke
13. Phosphoinositide Analysis by Liquid Chromatography-Mass Spectrometry
Trevor R. Pettitt
14. Imaging Phosphoinositide Dynamics in Living Cells
Anne Wuttke, Olof Idevall-Hagren, and Anders Tengholm
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