Cytochrome P450 Protocols

Cytochrome P450 Protocols

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For this second edition of their much praised Cytochrome P450, the editors have collected accounts of the essential core techniques that use the latest methodologies for the investigation of P450s. Highlights include protocols for spectral analysis and purification of P450s, enzymatic assays of P450s and flavin-containing monooxygenases (FMOs), expression of P450s and FMOs in heterologous systems, and the production and use of antipeptide antibodies. Additional chapters contain readily reproducible techniques for the transfection of hepatocytes for gene regulation studies, P450 reporter gene assays, in situ hybridization, and analysis of genetic polymorphisms. Although the emphasis is on P450s of mammalian origin, many of the readily reproducible methods described are suitable for P450s from any source.show more

Product details

  • Hardback | 364 pages
  • 181.4 x 230.6 x 19.1mm | 721.22g
  • Humana Press Inc.
  • Totowa, NJ, United States
  • English
  • Revised
  • 2nd ed. 2006
  • 47 black & white illustrations, biography
  • 1588294412
  • 9781588294418

Review quote

From Reviews of the First Edition... "...a remarkable book, highly useful for all our scientific readers..." -Cellular and Molecular Biology "...will be welcomed by laboratory scientists of various profiles who need authoritative protocols for their studies." -Modern Pathology From the reviews of the second edition: "In this second edition, editors Phillips and Shephard have updated the text to reflect current advances and to include chapters on some more contemporary procedures ... . I'm sure that the wealth of background information in the book would be valuable. Overall ... this is a useful reference book for experienced researchers and for those new to this discipline. Recommended." (David Wilkinson, British Toxicology Society Newsletter, Winter, 2006)show more

Back cover copy

For this second edition of their much praised Cytochrome P450, the editors have collected accounts of the essential core techniques that use the latest methodologies for the investigation of P450s. Highlights include protocols for spectral analysis and purification of P450s, enzymatic assays of P450s and flavin-containing monooxygenases (FMOs), expression of P450s and FMOs in heterologous systems, and the production and use of antipeptide antibodies. Additional chapters contain readily reproducible techniques for the transfection of hepatocytes for gene regulation studies, P450 reporter gene assays, in situ hybridization, and analysis of genetic polymorphisms. Also described are techniques for the generation of mice with targeted gene disruptions. Although the emphasis is on P450s of mammalian origin, many of the readily reproducible methods described are suitable for P450s from any source. Each chapter is written by researchers who have been involved in the development and application of the particular technique to P450s or FMOs. The protocols are presented in a step-by-step manner, with extensive cross-references to notes that highlight critical steps, potential problems, and alternative methods so that the researcher can understand the basis of the method and perform it successfully. Cutting-edge and highly practical, Cytochrome P450, Second Edition provides both novices and experienced researchers across many fields all the tools needed today to elucidate the crucial biological role played by cytochrome P450 in the metabolism of therapeutic drugs, chemical carcinogens, and environmental pollutants.show more

Table of contents

Cytochrome P450 Nomenclature, 2004 David R. Nelson Spectral Analyses of Cytochromes P450 John B. Schenkman and Ingela Jansson Establishment of Functional Human Cytochrome P450 Monooxygenase Systems in Escherichia coli Michael P. Pritchard, Lesley McLaughlin, and Thomas Friedberg Purification of Cytochromes P450: Products of Bacterial Recombinant Expression Systems F. Peter Guengerich and Martha V. Martin Expression of Recombinant Flavin-Containing Monooxygenases in a Baculovirus/Insect Cell System Azara Janmohamed, Prapimpuk Thaunsukon, Elizabeth A. Shephard, and Ian R. Phillips Cytochrome P450 Reconstitution Systems Hiroshi Yamazaki and Tsutomu Shimada Catalytic Assays for Human Cytochrome P450: An Introduction Thomas K. H. Chang and David J. Waxman Enzymatic Analysis of cDNA-Expressed Human CYP1A1, CYP1A2, and CYP1B1 With 7-Ethoxyresorufin as Substrate Thomas K. H. Chang and David J. Waxman Spectrofluorometric Analysis of CYP2A6-Catalyzed Coumarin 7-Hydroxylation David J. Waxman and Thomas K. H. Chang Determination of CYP2B6 Component of 7-Ethoxy-4-Trifluoromethylcoumarin O-Deethylation Activity in Human Liver Microsomes Thomas K. H. Chang, Charles L. Crespi, and David J. Waxman High-Performance Liquid Chromatography Analysis of CYP2C8-Catalyzed Paclitaxel 6a-Hydroxylation Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman Determination of CYP2C9-Catalyzed Diclofenac 4'-Hydroxylation by High-Performance Liquid Chromatography Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman CYP2C19-Mediated (S)-Mephenytoin 4'-Hydroxylation Assayed by High-Performance Liquid Chromatography With Radiometric Detection Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman CYP2D6-Dependent Bufuralol 1'-Hydroxylation Assayed by Reverse-Phase Ion-Pair High-Performance Liquid Chromatography With Fluorescence Detection Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman Spectrophotometric Analysis of Human CYP2E1-Catalyzed p-Nitrophenol Hydroxylation Thomas K. H. Chang, Charles L. Crespi, and David J. Waxman Thin-Layer Chromatography Analysis of Human CYP3A-Catalyzed Testosterone 6b-Hydroxylation David J. Waxman and Thomas K. H. Chang Determination of CYP4A11-Catalyzed Lauric Acid 12-Hydroxylation by High-Performance Liquid Chromatography With Radiometric Detection Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman An Isocratic High-Performance Liquid Chromatography Assay for CYP7A1-Catalyzed Cholesterol 7a-Hydroxylation David J. Waxman and Thomas K. H. Chang Use of 7-Ethoxycoumarin to Monitor Multiple Enzymes in the Human CYP1, CYP2, and CYP3 Families David J. Waxman and Thomas K. H. Chang Benzydamine N-Oxygenation as a Measure of Flavin-Containing Monooxygenase Activity Catherine K. Yeung and Allan E. Rettie Prochiral Sulfoxidation as a Probe for Flavin-Containing Monooxygenases Catherine K. Yeung and Allan E. Rettie Targeting Antipeptide Antibodies Toward Cytochrome P450 Enzymes Robert J. Edwards The Human Cytochrome P450 Allele Nomenclature Committee Web Site: Submission Criteria, Procedures, and Objectives Sarah C. Sim and Magnus Ingelman-Sundberg Genotyping for Cytochrome P450 Polymorphisms Ann K. Daly, Barry P. King, and Julian B. S. Leathart Hepatocyte Cultures in Drug Metabolism and Toxicological Research and Testing Tamara Vanhaecke and Vera Rogiers Isolation of Rat Hepatocytes Peggy Papeleu, Tamara Vanhaecke, Tom Henkens, Greetje Elaut, Mathieu Vinken, Sarah Snykers, and Vera Rogiers Rat Hepatocyte Cultures: Conventional Monolayer and Cocultures With Rat Liver Epithelial Cells Tom Henkens,show more

Review Text

From Reviews of the First Edition... "...a remarkable book, highly useful for all our scientific readers..." -Cellular and Molecular Biology "...will be welcomed by laboratory scientists of various profiles who need authoritative protocols for their studies." -Modern Pathology From the reviews of the second edition: "In this second edition, editors Phillips and Shephard have updated the text to reflect current advances and to include chapters on some more contemporary procedures ... . I'm sure that the wealth of background information in the book would be valuable. Overall ... this is a useful reference book for experienced researchers and for those new to this discipline. Recommended." (David Wilkinson, British Toxicology Society Newsletter, Winter, 2006)show more