Methods In Molecular Biology Cytochrome P450 Protocols

In Cytochrome P450 Protocols, Ian Phillips and Elizabeth Shephard assemble a comprehensive collection of cutting-edge techniques 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 polymorphisms. 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 Cytochrome P450 Protocols includes in one volume both state-of-the-art and classic methods that have not been superseded but remain extremely useful. The collection 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 cytochromes P450 in the metabolism of therapeutic drugs, chemical carcinogens, and environmental pollutants.