Full Description
Since the discovery of the pharmacological and toxicological importance of inhibiting the cyclooxygenase (COX) enzymes by non-steroidal anti-inflammatory drugs (NSAIDs), much research has gone into the development of methods to study the biological functions of COX-1 and COX-2. In Cyclooxygenases: Methods and Protocols, experts and pioneers in the field present the most up-to-date in vitro and in vivo techniques routinely used in COX research. The volume delves into essential topics such as the purification, cloning, and expression of COX enzymes as well as in vitro assays aimed at determining the inhibitory potency of drugs on COX-1 and COX-2 activities, with chapters describing protocols used for the extraction and measurement of the prostanoids. This volume also describes in vivo disease models used to study the roles of COX-1 and COX-2 in gastrointestinal injury, inflammation, and pain. As a book in the highly successful Methods in Molecular Biology™ series, the protocols chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Cyclooxygenases: Methods and Protocols serves as an indispensable tool for all scientists seeking the treatment of inflammation, pain, fever, and other harmful conditions.
Contents
In Vitro Protocols for Studying Expression and Activity of Cyclooxygenase Enzymes and Characterisation of Their Inhibition Patterns.- In Vitro Cyclo-oxygenase Expression and Activity Protocols: Introduction to Part I.- Techniques Used to Study Regulation of Cyclooxygenase-2 Promoter Sites.- Purification of Recombinant Human COX-1 and COX-2.- Cloning and Expression of Cyclooxygenase-1 and Cyclooxygenase-2.- Expression of Cyclooxygenase Isoforms in the Baculovirus Expression System.- Peroxidase Active Site Activity Assay.- Study of Inhibitors of the PGH Synthases for which Potency Is Regulated by the Redox State of the Enzymes.- Different Methods for Testing Potential Cyclooxygenase-1 and Cyclooxygenase-2 Inhibitors.- In Vitro Cyclooxygenase Activity Assay in Tissue Homogenates.- Techniques Used to Characterize the Binding of Cyclooxygenase Inhibitors to the Cyclooxygenase Active Site.- In Vitro Assays for Cyclooxygenase Activity and Inhibitor Characterization.- Extraction and Measurement of Prostanoids and Isoprostanes.- Extraction and Measurement of Prostanoids and Isoprostanes: Introduction to Part II.- Prostanoid Extraction and Measurement.- Measurement of 8-Iso-Prostaglandin F2? in Biological Fluids as a Measure of Lipid Peroxidation.- In Vivo Models to Study Involvement of Cyclooxygenase Products in Health and Disease.- In Vivo Models to Study Cyclooxygenase Products in Health and Disease: Introduction to Part III.- Protocols to Assess the Gastrointestinal Side Effects Resulting from Inhibition of Cyclo-Oxygenase Isoforms.- Cyclooxygenase Enzymes and Their Products in the Carrageenan-Induced Pleurisy in Rats.- Iloprost-Induced Nociception: Determination of the Site of Anti-nociceptive Action of Cyclooxygenase Inhibitors and the Involvement of CyclooxygenaseProducts in Central Mechanisms of Nociception.
