Ethoxyresorufin-O-deethylase and porphyrin analysis in chicken embryo hepatocyte cultures with a fluorescence multiwell plate reader

Anal Biochem. 1993 May 15;211(1):102-12. doi: 10.1006/abio.1993.1239.

Abstract

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 in 48-well cell culture plates, and EROD assays were carried out directly within the wells. EROD activity and porphyrin concentrations were determined with a computer-operated fluorescence multiwell plate reader. Porphyrin and EROD assays could be carried out on the same plates or on separate plates. The assays allow one to obtain dose-response curves at rates that are approximately 100 times faster than conventional methods.

MeSH terms

  • Animals
  • Cells, Cultured
  • Chick Embryo
  • Chromatography, High Pressure Liquid
  • Computers
  • Cytochrome P-450 CYP1A1
  • Cytochrome P-450 Enzyme System / analysis*
  • Kinetics
  • Liver / chemistry*
  • Liver / cytology
  • Liver / enzymology
  • Oxidoreductases / analysis*
  • Porphyrins / analysis*
  • Reference Standards
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence / instrumentation

Substances

  • Porphyrins
  • Cytochrome P-450 Enzyme System
  • Oxidoreductases
  • Cytochrome P-450 CYP1A1