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Efficient Analysis of Cytochrome P4501A Catalytic Activity, Porphyrins, and Total Proteins in Chicken Embryo Hepatocyte Cultures with a Fluorescence Plate Reader

✍ Scribed by S.W. Kennedy; S.P. Jones; L.J. Bastien

Book ID: 102968614
Publisher: Elsevier Science
Year: 1995
Tongue: English
Weight: 576 KB
Volume: 226
Category: Article
ISSN: 0003-2697
DOI: 10.1006/abio.1995.1237

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✦ Synopsis

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. (\Leftrightarrow, 1995) Academic Press, Inc.

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Rapid and sensitive methods for measuring ethoxyresorufin-O-deethylase (EROD) induction and total intracellular porphyrin accumulation in cultured chicken embryo hepatocytes exposed to polychlorinated biphenyls (PCBs) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are described. Cells were cultured