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Inositol Phosphates and Lipids Methods and Protocols




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Dettagli

Genere:Libro
Lingua: Inglese
Editore:

Humana Press

Edizione: 2010





Trama

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.





Sommario

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 InsP 7 by the Inositol Hexakisphosphate Kinase 1 (IP6K1).- Protein Pyrophosphorylation by Diphosphoinositol Pentakisphosphate (InsP 7).- 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 Interference.- Phosphoinositide-Specific Phospholipase C ?1 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.










Altre Informazioni

ISBN:

9781603271745

Condizione: Nuovo
Collana: Methods in Molecular Biology
Dimensioni: 254 x 178 mm
Formato: Copertina rigida
Illustration Notes:X, 230 p. 54 illus.
Pagine Arabe: 230
Pagine Romane: x


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