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Phospholipid Signaling Protocols

Series: Methods in Molecular Biology

by Ian Bird

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Main description:

Cell membranes are not, as once believed, inert structures designed to contain the cell contents, but are in fact dynamic structures that are as me- bolically active as the cytosol and other cellular compartments they surround. Thus membranes not only contain mixtures of lipid and phospholipids, but also many proteins both embedded deeply within the membrane structure itself and also more loosely attached on the membrane surfaces. Though many such proteins have long been known to act as transport proteins, ion channels, hormone receptors, G proteins, cytoskeletal anchorage points, and so on, the major advance of recent years is the increasing understanding that the lipids and phospholipids in the membrane bilayer itself are also metabolized to b- logically active products that can diffuse either in the cytosol or in the m- brane bilayer to control the function of other proteins. Thus the concept of lipid-derived second messengers is now firmly established.

Contents:

Part I. Monitoring of Activation of Phospholipid-Derived Cell Signaling Pathways

Phosphoinositidase C Activation Assay I: Cell Labeling, Stimulation, and Recovery of Cellular [3H] Phosphoinositides and [3H] Phosphoinositols
Ian M. Bird

Phosphoinositidase C Activation Assay II: Simple Analysis of Recovered Cellular Phosphoinositides and Phosphoinositols
Ian M. Bird

Phosphoinositidase C Activation Assay III: HPLC Analysis of Cellular Phosphoinositides and Phosphoinositols
Ian M. Bird

Measurement of Phosphoinositols and Phosphoinositides Using Radio High-Performance Liquid Chromatography Flow Detection
Lubo Zhang and Iain L. O. Buxton

Preparation of [3H]Phosphoinositol Standards and Conversion of [3H] Phosphoinositides to [3H] Phosphoinositols
Ian M. Bird

Inositol 1,4,5-Trisphosphate Mass Assay
Lubo Zhang

Measurement of Cellular Diacylglycerol Content
Wendy Bollinger Bollag and Richard D. Griner

Phosphatidylinositol 4-Kinases: Assays and Product Analysis
Rachel E. Meyers and Lewis C. Cantley

Phosphatidylinositol(3,4,5)Trisphosphate (Ptdins(3,4,5)P3) Mass Measurement Using a Radioligand Displacement Assay
Jeroen van der Kaay, Pete J. Cullen, and C. Peter Downes

Detection of Phosphatidylinositol-4-Phosphate 5-Kinase Activity Using Thin-Layer Chromatography
Gregory J. Parker, Joost C. Loijens, and Richard A. Anderson

Determination of Phospholipase C- or Phospholipase D-Catalyzed Phosphatidylcholine Hydrolysis
Michael J. O. Wakelam and Trevor R. Pettitt

Measurement of Phospholipase D Activity
Wendy Bollinger Bollag

Monitoring of Phospholipase A2 Activation in Cultured Cells Using Tritiated Arachidonic Acid
Sandie I. Briand, Sylvie G. Bernier, and Gaetan Guillemette

Assay of Cellular Diacylglycerol and Monoacylglycerol Lipases
Bryan A.Wolf

Isotopic Efflux Studies as Indices of Phospholipase Activation
Jacob Vadakekalam and Stewart Metz

Extraction and Measurements of Prostanoids and Leukotrienes by Radioimmunoassays
Bruno Battistini, Serge Picard, Pierre Borgeat, and Pierre Sirois

Measurement of Prostanoids, Leukotrienes, and Isoprostanes By Enzyme Immunoassays
Bruno Battistini, Serge Picard, Pierre Borgeat, and Pierre Sirois

Measurements of Arachidonic Acid Metabolites Derived from the Lipoxygenase Pathways By High-Pressure Liquid Chromatography
Pierre Borgeat, Serge Picard, Bruno Battistini, and Pierre Sirois

Measurement of Sphingomyelin and Ceramide Cellular Levels After Sphingomyelinase-Mediated Sphingomyelin Hydrolysis
Pino Santana, Luisa F. Fanjul, and C. M. Ruiz de Galarreta

Sphingomyelin and Ceramide Mass Assay
Pino Santana, C. M. Ruiz de Galarreta, and Luisa F. Fanjul

Sphingosine Kinase: Assay and Product Analysis
Ana Olivera and Sarah Spiegel

Analytical Methods and Steps to Sample Preparation For Determination of Molecular Species of Fatty Acids
Sanda Clejan

HPLC Analytical Methods for the Separation of Molecular Species of Fatty Acids In Diacylglycerol and Cellular Phospholipids
Sanda Clejan

Analysis of Molecular Species of Cellular Sphingomyelins and Ceramides
Sanda Clejan

Part II. General methods for monitoring changes in levels of key signaling pathway proteins and associated mRNA

Solubilization and Assay of Cellular and Tissue Protein
Cynthia E. Shaw

Western Immunoblot Analysis
Cynthia E. Shaw and Jing Zheng

Immunohistochemistry and Immunocytochemistry
Jing Zheng

Extraction of Cellular and Tissue RNA
Daniel S. Millican and Ian M. Bird

Size Separation and Quantification of mRNA by Northern Analysis
Ian M. Bird

Preparation of

PRODUCT DETAILS

ISBN-13: 9781489942609
Publisher: Springer (Humana Press Inc.)
Publication date: August, 2013
Pages: 380
Weight: 584g
Availability: Available
Subcategories: Biochemistry

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