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This readily reproducible collection of cutting-edge methods for the study of eicosanoids concentrates on studying the regulation of expression and function of enzymes, particularly cyclooxygenase (and its two isoforms), phospholipase A2, and lipoxygenases involved in the synthesis of established eicosanoids. Additional chapters are devoted to the characterization and distribution of the thromboxane A2 receptor in tissues and the biological roles of novel eicosanoids. Eicosanoid Protocols offers new and established researchers powerful, state-of-the-art tools to probe the regulation and function of eicosanoids.
Contents
Assessment of Arachidonic Acid Distribution into Phospholipids of Inflammatory Cells Alfred N. Fonteh. Determination of Phospholipase A2s Expression in Mast Cells by Reverse- Transcriptase-Polymerase Chain Reaction Alfred N. Fonteh William Reed and James M. Samet. Extraction of Eicosanoids from Biological Fluids Cells and Tissues William S. Powell. Separation of Hydroxyeicosatetraenoic Acids and Leukotrienes Originating from Kidney Tissue by C-18 Reverse- phase High-pressure Liquid Chromatography Mukut Sharma Beatrice Hucke and Elias A. Lianos. Enzyme Immunoassays (EIA) of Eicosanoids Kingsley Yin and Lloyd J. Forman. Assessment of Cyclooxygenase RNA Expression by Northern Hybridization Zhonghong Guan and Aubrey R. Morrison. Assessment of Cyclooxygenase Protein Expression by Western Blotting Zhonghong Guan and Aubrey R. Morrison. Use of Retroviral Vector for Expression of a Foreign Gene Brent W. Miller and Aubrey R. Morrison. Assessment of the Expression of Prostaglandin Synthase-2 in Swiss 3T3 Fibroblasts Srinivasa T. Reddy and Harvey R. Herschman. Localization and Distribution of Cyclooxygenase-2 in Brain Tissue by Immunohistochemistry M. Kerry O'Banion and John A. Olschowka. Immunohistochemical Analysis of Cyclooxygenase Expression in Human Skin ShaAvhree Y. Buckman Alane T. Koki Dorothy A. Edwards and Alice P. Pentland. Assessment of 5-Lipoxygenase Activity and Cellular Distribution Robert A. Lepley. Whole Blood Assay for Evaluation of Thromboxane Synthase Inhibition Eric G. Spokas Nancy S. Nicholson Osman D. Sulemanov Gerald M. Walsh and David Tuffin. Assessment of Cellular Localization of the Thromboxane A2 Receptor by Immunocytochemistry Samuel C. Blackman Chatherine Borg David G. Yeomans and Guy C. LeBreton. Transcellular Regulation of Eicosanoid Biosynthesis Karsten Gronert Clary B. Clish Mario Romano and Charles N. Serhan. Leukotrienes Lipoxins and Hydroxyeicosatetraenoic Acids Eric G. Spokas Joshua Rokach and Patrick Y-K Wong. Epoxyeicosatrienoic Acids: Biosynthesis Regulation and Actions John D. Imig. Novel Eicosanoids: Isoprostanes and Related Compounds L. Jackson Roberts II Cynthia J. Branes Yan Chan and Jason D. Morron.
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