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Cytochrome P450 Protocols

Ian R. Phillips

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Description:Ian Phillips and Elizabeth Shephard assemble a comprehensive collection of cutting-edge techniques and invaluable classic methods for the investigation of cytochromes P450. Described in detail by hands-on experimentalists for easy reproducibility, these methods include spectral analysis, purification and enzymatic assays, expression in heterologous systems, and the production and use of antibodies, as well as methods for quantification of gene expression, transfection of hepatocytes, and for the investigation of DNA protein interactions and genetic polymorphism. In addition, because of the growing importance of in vitro systems in pharmacological toxicology, the book contains techniques for the culture of rodent and human hepatocytes and human epidermis. Cytochrome P450 induction as a biomarker for environmental pollution and the generation of mice with targeted gene disruptions complete this exhaustive collection of core techniques.; Cytochrome P450 Protocols provides both novice and experienced researchers across many fields - toxicology, pharmacology, environmental biology, biochemistry, and molecular biology - all the tools needed to elucidate the crucial biological role played by the cytochromes P450 in the metabolism of therapeutic drugs, chemical carcinogens, and environmental pollutants.Contents: 1 The Role of Methods in the Discovery of the Cytochromes P450 2 Cytochrome P450 Nomenclature 3 Spectral Analyses of Cytochromes P450 4 Purification of Cytochromes P450: Rat and Human Hepatic Forms 5 Isolation and Purification of Constitutive Forms of Microsomal Cytochrome P450 6 Purification of Extrahepatic Cytochromes P450 7 Purification of Cytochromes P450: Products of Bacterial Recombinant Expression Systems 8 Cytochrome P450 Reconstitution Systems 9 Catalytic Assays for Human Cytochrome P450 10 Enzymatic Analysis of cDNA-Expressed Human CYP1A1, CYP1A2, and CYP1B1 with 7-Ethoxyresorufin as Substrate 11 Spectrofluorometric Analysis of CYP2A6-Catalyzed Coumarin 7-Hydroxylation 12 Determination of the CYP2B6 Component of 7-Ethoxy-4- Trifluoromethylcoumarin O-Deethylation Activity in Human Liver Microsomes 13 High-Performance Liquid Chromatographic Analysis of CYP2C8-Catalyzed Paclitaxel 6α-Hydroxylation 14 Determination of CYP2C9-Catalyzed Diciofenac 4'-Hydroxylation by High-Performance Liquid Chromatography 15 CYP2C19-Mediated (S)-Mephenytoin 4'-Hydroxylation Assayed by High-Performance Liquid Chromatography with Radiometric Detection 16 CYP2D6-Dependent Bufuralol 1'-Hydroxylation Assayed by Reversed-Phase Ion-Pair High-Performance Liquid Chromatography with Fluorescence Detection 17 Spectrophotometric Analysis of Human CYP2E1-Catalyzed p-Nitrophenol Hydroxylation 18 Thin-Layer Chromatographic Analysis of Human CYP3A-Catalyzed Testosterone 6β-Hydroxylation 19 Determination of CYP4A11-Catalyzed Lauric Acid 12-Hydroxylation by High-Performance Liquid Chromatography with Radiometric Detection 20 An Isocratic High-Performance Liquid Chromatographic Assay for CYP7A1-Catalyzed Cholesterol 7α-Hydroxylation 21 Use of 7-Ethoxycoumarin to Monitor Multiple Enzymes in the Human CYP1, CYP2, and CYP3 Families 22 Expression of Eukaryotic Cytochromes P450 in E. Coli 23 Expression of Mammalian Cytochromes P450 in Yeast 24 Expression of Cytochromes P450 in a Baculovirus System 25 Expression of Cytochromes P450 in Mammalian Cells 26 Monoclonal Antibodies to Cytochromes P450 27 Targeting Antipeptide Antibodies Towards Cytochrome P450 Enzymes 28 Signals for Retention of Cytochrome P450 in the Endoplasmic Reticulum 29 Quantification of Cytochrome P450 Gene Expression by RNase Protection Analysis 30 Hepatocyte Cultures in Drug Metabolism and Toxicological Research and Testing 31 Isolation of Rat Hepatocytes 32 Collagen-Gel Cultures of Rat Hepatocytes: Collagen-Gel Sandwich and Immobilization Cultures 33 Rat Hepatocyte Cultures: Conventional Monolayer Cultures and Cocultures with Rat Liver Epithelial Cells 34 Isolation of Periportal and Pericentral Hepatocytes 35 Perifusion Culture of Hepatocytes 36 Human Hepatocyte Culture 37 Reconstruction of Human Epidermis in Vitro 38 Gene Transfer and Expression 39 Lipid-Mediated Gene Transfer into Normal Adult Human Hepatocytes in Primary Culture 40 Identification of Regulatory Protein-Binding Sites in Cytochrome P450 Genes by Means of the Gel-Retardation Assay 41 Cytochrome P450 Gene Regulation: Analysis of Protein-DNA Interactions In Situ 42 Analysis of Cytochrome P450 Polymorphisms 43 Induction of Cytochrome P450 1A (CYP1A) in Fish: A Biomarker for Environmental Pollution 44 Generation of Allelic Series in Mice by Gene TargetingWe have made it easy for you to find a PDF Ebooks without any digging. And by having access to our ebooks online or by storing it on your computer, you have convenient answers with Cytochrome P450 Protocols. To get started finding Cytochrome P450 Protocols, you are right to find our website which has a comprehensive collection of manuals listed.
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ISBN: 1592599982

Description: Ian Phillips and Elizabeth Shephard assemble a comprehensive collection of cutting-edge techniques and invaluable classic methods for the investigation of cytochromes P450. Described in detail by hands-on experimentalists for easy reproducibility, these methods include spectral analysis, purification and enzymatic assays, expression in heterologous systems, and the production and use of antibodies, as well as methods for quantification of gene expression, transfection of hepatocytes, and for the investigation of DNA protein interactions and genetic polymorphism. In addition, because of the growing importance of in vitro systems in pharmacological toxicology, the book contains techniques for the culture of rodent and human hepatocytes and human epidermis. Cytochrome P450 induction as a biomarker for environmental pollution and the generation of mice with targeted gene disruptions complete this exhaustive collection of core techniques.; Cytochrome P450 Protocols provides both novice and experienced researchers across many fields - toxicology, pharmacology, environmental biology, biochemistry, and molecular biology - all the tools needed to elucidate the crucial biological role played by the cytochromes P450 in the metabolism of therapeutic drugs, chemical carcinogens, and environmental pollutants.Contents: 1 The Role of Methods in the Discovery of the Cytochromes P450 2 Cytochrome P450 Nomenclature 3 Spectral Analyses of Cytochromes P450 4 Purification of Cytochromes P450: Rat and Human Hepatic Forms 5 Isolation and Purification of Constitutive Forms of Microsomal Cytochrome P450 6 Purification of Extrahepatic Cytochromes P450 7 Purification of Cytochromes P450: Products of Bacterial Recombinant Expression Systems 8 Cytochrome P450 Reconstitution Systems 9 Catalytic Assays for Human Cytochrome P450 10 Enzymatic Analysis of cDNA-Expressed Human CYP1A1, CYP1A2, and CYP1B1 with 7-Ethoxyresorufin as Substrate 11 Spectrofluorometric Analysis of CYP2A6-Catalyzed Coumarin 7-Hydroxylation 12 Determination of the CYP2B6 Component of 7-Ethoxy-4- Trifluoromethylcoumarin O-Deethylation Activity in Human Liver Microsomes 13 High-Performance Liquid Chromatographic Analysis of CYP2C8-Catalyzed Paclitaxel 6α-Hydroxylation 14 Determination of CYP2C9-Catalyzed Diciofenac 4'-Hydroxylation by High-Performance Liquid Chromatography 15 CYP2C19-Mediated (S)-Mephenytoin 4'-Hydroxylation Assayed by High-Performance Liquid Chromatography with Radiometric Detection 16 CYP2D6-Dependent Bufuralol 1'-Hydroxylation Assayed by Reversed-Phase Ion-Pair High-Performance Liquid Chromatography with Fluorescence Detection 17 Spectrophotometric Analysis of Human CYP2E1-Catalyzed p-Nitrophenol Hydroxylation 18 Thin-Layer Chromatographic Analysis of Human CYP3A-Catalyzed Testosterone 6β-Hydroxylation 19 Determination of CYP4A11-Catalyzed Lauric Acid 12-Hydroxylation by High-Performance Liquid Chromatography with Radiometric Detection 20 An Isocratic High-Performance Liquid Chromatographic Assay for CYP7A1-Catalyzed Cholesterol 7α-Hydroxylation 21 Use of 7-Ethoxycoumarin to Monitor Multiple Enzymes in the Human CYP1, CYP2, and CYP3 Families 22 Expression of Eukaryotic Cytochromes P450 in E. Coli 23 Expression of Mammalian Cytochromes P450 in Yeast 24 Expression of Cytochromes P450 in a Baculovirus System 25 Expression of Cytochromes P450 in Mammalian Cells 26 Monoclonal Antibodies to Cytochromes P450 27 Targeting Antipeptide Antibodies Towards Cytochrome P450 Enzymes 28 Signals for Retention of Cytochrome P450 in the Endoplasmic Reticulum 29 Quantification of Cytochrome P450 Gene Expression by RNase Protection Analysis 30 Hepatocyte Cultures in Drug Metabolism and Toxicological Research and Testing 31 Isolation of Rat Hepatocytes 32 Collagen-Gel Cultures of Rat Hepatocytes: Collagen-Gel Sandwich and Immobilization Cultures 33 Rat Hepatocyte Cultures: Conventional Monolayer Cultures and Cocultures with Rat Liver Epithelial Cells 34 Isolation of Periportal and Pericentral Hepatocytes 35 Perifusion Culture of Hepatocytes 36 Human Hepatocyte Culture 37 Reconstruction of Human Epidermis in Vitro 38 Gene Transfer and Expression 39 Lipid-Mediated Gene Transfer into Normal Adult Human Hepatocytes in Primary Culture 40 Identification of Regulatory Protein-Binding Sites in Cytochrome P450 Genes by Means of the Gel-Retardation Assay 41 Cytochrome P450 Gene Regulation: Analysis of Protein-DNA Interactions In Situ 42 Analysis of Cytochrome P450 Polymorphisms 43 Induction of Cytochrome P450 1A (CYP1A) in Fish: A Biomarker for Environmental Pollution 44 Generation of Allelic Series in Mice by Gene TargetingWe have made it easy for you to find a PDF Ebooks without any digging. And by having access to our ebooks online or by storing it on your computer, you have convenient answers with Cytochrome P450 Protocols. To get started finding Cytochrome P450 Protocols, you are right to find our website which has a comprehensive collection of manuals listed.
Our library is the biggest of these that have literally hundreds of thousands of different products represented.

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