Abstract
An efficient method for measuring polychlorinated biphenyl-mediated induction of cytochrome P4501A, using the ethoxyresorufin-O-deethylase (EROD) assay, and total porphyrins in chicken embryo hepatocyte cultures is described. Hepatocytes were cultured in 48-well plates, assays were carried out within the wells, and concentrations of the product of the EROD reaction (resorufin), porphyrins, and total proteins were measured in the same wells with a fluorescence plate reader. The method differs from previous methods developed in this laboratory in that three fluorophors were measured in the same well rather than two.
MeSH terms
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Animals
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Cells, Cultured
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Chick Embryo
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Chromatography, High Pressure Liquid
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Cytochrome P-450 CYP1A1
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Cytochrome P-450 Enzyme System / metabolism*
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Fluorescamine
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Hydrochloric Acid / pharmacology
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Liver / chemistry
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Liver / cytology
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Liver / metabolism*
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Oxazines / analysis
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Oxazines / metabolism
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Oxidoreductases / metabolism*
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Polychlorinated Biphenyls / pharmacology
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Porphyrins / analysis*
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Porphyrins / metabolism
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Proteins / analysis*
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Proteins / metabolism
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Spectrometry, Fluorescence
Substances
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Oxazines
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Porphyrins
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Proteins
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Fluorescamine
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ethoxyresorufin
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resorufin
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Cytochrome P-450 Enzyme System
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Polychlorinated Biphenyls
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Oxidoreductases
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Cytochrome P-450 CYP1A1
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Hydrochloric Acid