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 BiologyT 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.
Part I: In vitro Protocols for Studying Expression and Activity of Cyclooxygenase Enzymes and Characterisation of Their Inhibition Patterns 1. In vitro Expression and Activity Protocols: Introduction to Part I Roderick J. Flower 2. Techniques Used to Study Regulation of Cyclooxygenase-2 Promoter Sites Hiroyasu Inoue and Rieko Nakata 3. Purification of Recombinant Human COX-1 and COX-2 James K. Gierse 4. Cloning and Expression of Cyclooxygenase-1 and Cyclooxygenase-2 Nicholas R. Staten and Beverly A. Reitz 5. Expression of Cyclooxygenase Isoforms in the Baculovirus Expression System John C. Hunter, Natalie M. Myres, and Daniel L. Simmons 6. Peroxidase Active Site Activity Assay Kelsey C. Duggan, Joel Musee, and Lawrence J. Marnett 7. Study of Inhibitors of the PGH Synthases for which Potency is Regulated by the Redox State of the Enzymes Olivier Boutaud and John A. Oates 8. Different Methods for Testing Potential Cyclooxygenase-1 and Cyclooxygenase-2 Inhibitors Stefan Laufer and Sabine Luik 9. In vitro Cyclooxygenase Activity Assay in Tissue Homogenates Samir S. Ayoub 10. Techniques Used to Characterize the Binding of Cyclooxygenase Inhibitors to the Cyclooxygenase Active Site William F. Hood 11. In vitro Assays for Cyclooxygenase Activity and Inhibitor Characterization Mark C. Walker and James K. Gierse Part II: Extraction and Measurement of Prostanoids and Isoprostanes 12. Extraction and Measurement of Prostanoids and Isoprostanes: Introduction to Part II Paola Patrignani 13. Prostanoid Extraction and Measurement Lorenzo Polenzani and Samir S. Ayoub 14. Measurement of 8-iso-prostagnadin F2a in Biological Fluids as a Measure of Lipid Peroxidation Stefania Tacconelli, Marta L. Capone, and Paola Patrignani Part III: In vivo Models to Study Involvement of Cyclooxygenase Products in Health and Disease 15. In vivo Models to Study Cyclooxygenase Products in Health and Disease: Introduction to Part III Derek W. Gilroy, Melanie Stables, and Justine Newson 16. Protocols to Assess the Gastro-Intestinal Side Effects Resulting from Inhibition of Cyclo-Oxygenase Isoforms Brendan J.R. Whittle 17. Cyclooxygenase Enzymes and Their Products in the Carragenaan-Induced Pleurisy in Rats Adrian R. Moore, Samir S. Ayoub, and Michael P. Seed 18. Iloprost-Induced Nociception: Determination of the Site of Anti-Nociceptive Action of Cyclooxygenase Inhibitors and the Involvement of Cyclooxygenase Products in Central Mechanisms of Nociception Samir S. Ayoub and Regina M. Botting
Series: Methods in Molecular Biology
Number Of Pages: 218
Published: 7th May 2010
Publisher: Humana Press Inc.
Country of Publication: US
Dimensions (cm): 25.4 x 17.8
Weight (kg): 1.37