Simultaneous measurement of cytochrome P4501A catalytic activity and total protein concentration with a fluorescence plate reader.
A method for simultaneous measurement of the cytochrome P4501A-associated enzyme, ethoxyresorufin-O-deethylase (EROD), and total protein concentration in liver microsomes is described. EROD assays were carried out in multiwell plates, and the fluorescent product (resorufin) and total proteins were quantified within the same wells with a fluorescence plate reader. Fluorescamine was used for protein assays.
Canadian Wildlife Service, National Wildlife Research Centre, Hull, Quebec.
S W
SW
Kennedy
S P
SP
Jones
D000818:Animals; D002384:Catalysis; D019363:Cytochrome P-450 CYP1A1; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis*; D005450:Fluorescamine; D005990:Glycerol / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology; D009249:NADP / Q000494:pharmacology; D010078:Oxazines / Q000032:analysis; D010088:Oxidoreductases / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013050:Spectrometry, Fluorescence
D007527:Isoenzymes; D010078:Oxazines; D011506:Proteins; D005450:Fluorescamine; D009249:NADP; C014180:resorufin; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D019363:Cytochrome P-450 CYP1A1; D005990:Glycerol