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Cytochrome P450 Protocols pp 267–277Cite as

Quantification of Cytochrome P450 Gene Expression by RNase Protection Analysis

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 107))

Abstract

Many attempts to describe the regulation of mdivldual cytochrome P450 (CYP) gene products in animal tissues have been frustrated by the lack of sensitivity and specificity of the assays used. Most enzymatic assays are hampered by the overlapping substrate specificity of these enzymes, and the generation of high-affinity antibodies that are specific for individual isoforms is not a trivial task. Many CYP isoforms have close relatives, some of which can have as much as 98% identity at the RNA nucleotlde-sequence level, and filter-hybridization techniques using labeled cDNA probes are unable to dlscrlminate between such mRNAs. Although noncoding portions of the cDNAs may be used as probes, as they are more likely to offer regions of divergence within subfamllles, this is not often possible because

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Author information

Authors and Affiliations

  1. Biomedical Research Center, Ninewells Hospital and Medical School, University of Dundee, Scotland, UK

    Colin N. A. Palmer

Authors
  1. Colin N. A. Palmer
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Editor information

Editors and Affiliations

  1. Department of Biochemistry, Queen Mary and Westfie1d College, University of London, UK

    Ian R. Phillips

  2. Department of Biochemistry and Molecular Biology, University College, London, UK

    Elizabeth A. Shephard

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© 1998 Humana Press Inc.

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N. A. Palmer, C. (1998). Quantification of Cytochrome P450 Gene Expression by RNase Protection Analysis. In: Phillips, I.R., Shephard, E.A. (eds) Cytochrome P450 Protocols. Methods in Molecular Biology™, vol 107. Humana Press. https://doi.org/10.1385/0-89603-519-0:267

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  • DOI: https://doi.org/10.1385/0-89603-519-0:267

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-519-5

  • Online ISBN: 978-1-59259-580-8

  • eBook Packages: Springer Protocols

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