Cytochrome P450 Protocols / Edition 2

Cytochrome P450 Protocols / Edition 2

by Ian R. Phillips
     
 

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ISBN-10: 1617375721

ISBN-13: 9781617375729

Pub. Date: 11/19/2010

Publisher: Springer-Verlag New York, LLC

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

Overview

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.

Product Details

ISBN-13:
9781617375729
Publisher:
Springer-Verlag New York, LLC
Publication date:
11/19/2010
Series:
Methods in Molecular Biology Series, #320
Edition description:
2nd ed. 2006
Pages:
364
Product dimensions:
6.10(w) x 9.25(h) x 0.03(d)

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, Tamara Vanhaecke, Peggy Papeleu, Greetje Elaut, Mathieu Vinken, Sarah Snykers, and Vera Rogiers

Rat Hepatocyte Cultures: Collagen Gel Sandwich and Immobilization Cultures

Mathieu Vinken, Greetje Elaut, Tom Henkens, Peggy Papeleu, Sarah Snykers, Tamara Vanhaecke, and Vera Rogiers

Hepatocytes in Suspension

Greetje Elaut, Peggy Papeleu, Mathieu Vinken, Tom Henkens, Sarah Snykers, Tamara Vanhaecke, and Vera Rogiers

Isolation of Rat Bone Marrow Stem Cells

Sarah Snykers, Tamara Vanhaecke, and Vera Rogiers

Transfection of Primary Cultures of Rat Hepatocytes

Mina Edwards, Siew Cheng Wong, Roongsiri Chotpadiwetkul, Despina Smirlis, Ian R. Phillips, and Elizabeth A. Shephard

Human Hepatocyte Culture

Lydiane Pichard, Edith Raulet, Gérard Fabre, Jean Bernard Ferrini, Jean-Claude Ourlin, and Patrick Maurel

Determination of Cellular Localization of the Expression of Flavin-Containing Monooxygenase Genes in Mouse Tissues by In Situ Hybridization

Azara Janmohamed, Ian R. Phillips, and Elizabeth A. Shephard

Deletion of Genes From the Mouse Genome Using Cre/loxP Technology

Diana Hernandez, Pritpal Chandan, Azara Janmohamed, Ian R. Phillips, and Elizabeth A. Shephard

Characterization of Targeted Mouse Embryonic Stem Cell Chromosomes: Karyotyping and Fluorescence In Situ Hybridization of Metaphase Spreads

Diana Hernandez, Ian R. Phillips, and Elizabeth A. Shephard

Microinjection of Targeted Embryonic Stem Cells and Establishment of Knockout Mouse Lines for Fmo Genes

Diana Hernandez, Anna N. Melidoni, Ian R. Phillips, and Elizabeth A. Shephard

Use of Reporter Genes to Measure Xenobiotic-Mediated Activation of CYP Gene Transcription

Nick Plant

Index

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