Efficient analysis of cytochrome P4501A catalytic activity, porphyrins, and total proteins in chicken embryo hepatocyte cultures with a fluorescence plate reader.
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.
Kennedy
S W
SW
Canadian Wildlife Service, National Wildlife Research Centre, Hull, Quebec.
Jones
S P
SP
Bastien
L J
LJ
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002851:Chromatography, High Pressure Liquid; D019363:Cytochrome P-450 CYP1A1; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005450:Fluorescamine; D006851:Hydrochloric Acid / Q000494:pharmacology; D008099:Liver / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D010078:Oxazines / Q000032:analysis / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism*; D011078:Polychlorinated Biphenyls / Q000494:pharmacology; D011166:Porphyrins / Q000032:analysis* / Q000378:metabolism; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D010078:Oxazines; D011166:Porphyrins; D011506:Proteins; D005450:Fluorescamine; C007649:ethoxyresorufin; C014180:resorufin; D003577:Cytochrome P-450 Enzyme System; D011078:Polychlorinated Biphenyls; D010088:Oxidoreductases; D019363:Cytochrome P-450 CYP1A1; D006851:Hydrochloric Acid