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E-grāmata: Inositol Phosphates and Lipids: Methods and Protocols

  • Formāts: EPUB+DRM
  • Sērija : Methods in Molecular Biology 645
  • Izdošanas datums: 20-Jul-2010
  • Izdevniecība: Humana Press Inc.
  • Valoda: eng
  • ISBN-13: 9781603271752
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  • Formāts: EPUB+DRM
  • Sērija : Methods in Molecular Biology 645
  • Izdošanas datums: 20-Jul-2010
  • Izdevniecība: Humana Press Inc.
  • Valoda: eng
  • ISBN-13: 9781603271752
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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 v
Contributors ix
1 The Role of Inositol and the Principles of Labelling, Extraction, and Analysis of Inositides in Mammalian Cells
1(20)
Christopher J. Barker
Per-Olof Berggren
2 HPLC Separation of Inositol Polyphosphates
21(26)
Christopher J. Barker
Christopher Illies
Per-Olof Berggren
3 Molecular Manipulation and Analysis of Inositol Phosphate and Pyrophosphate Levels in Mammalian Cells
47(14)
James C. Otto
John D. York
4 A Femtomole-Sensitivity Mass Assay for Inositol Hexakisphosphate
61(12)
Andrew J. Letcher
Michael J. Schell
Robin F. Irvine
5 Synthesis of InsP, by the Inositol Hexakisphosphate Kinase I (IP6KI)
73(14)
Cristina Azevedo
Adam Burton
Matthew Bennett
Sara Maria Nancy Onnebo
Adolfo Saiardi
6 Protein Pyrophosphorylation by Diphosphoinositol Pentakisphosphate (InsP7)
87(16)
J. Kent Werner Jr.
Traci Speed
Rashna Bhandari
7 Synthesis and Nonradioactive Micro-analysis of Diphosphoinositol Phosphates by HPLC with Postcolumn Complexometry
103(20)
Hongying Lin
Karsten Lindner
Georg W. Mayr
8 Diphosphosinositol Polyphosphates and Energy Metabolism: Assay for ATP/ADP Ratio
123(10)
Andreas Nagel
Christopher J. Barker
Per-Olof Berggren
Christopher Illies
9 Isolation of Inositol 1, 4, 5-Trisphosphate Receptor-Associating Proteins and Selective Knockdown Using RNA Interference
133(10)
Akihiro Mizutani
Katsuhiro Kawaai
Chihiro Hisatsune
Hideaki Ando
Takayuki Michikawa
Katsuhiko Mikoshiba
10 Phosphoinositide-Specific Phospholipase C β l Signal Transduction in the Nucleus
143(22)
Roberta Fiume
Gabriella Teti
Irene Faenza
Lucio Cocco
11 Methods to Assess Changes in the Pattern of Nuclear Phosphoinositides
165(14)
Nullin Divecha
12 Measurement of Phosphoinositide 3-Kinase Products in Cultured Mammalian Cells by HPLC
179(24)
Frank T. Cooke
13 Phosphoinositide Analysis by Liquid Chromatography-Mass Spectrometry
203(16)
Trevor R. Pettitt
14 Imaging Phosphoinositide Dynamics in Living Cells
219(18)
Anne Wuttke
Olof Idevall-Hagren
Anders Tengholm
Index 237