PURIFICATION AND IMMUNOCHEMICAL DETECTION OF BETA-NAPHTHOFLAVONE-INDUCED AND PHENOBARBITAL-INDUCED AVIAN CYTOCHROME-P450 ENZYMES

Livers from mallards (Anas platyrhynchos) were treated with either bet a-naphthoflavone (50 mg/kg) or phenobarbital (70 mg/kg). Purification of induced hepatic cytochrome P450 was accomplished using both DEAF an d hydroxyapatite columns, as well as sodium dodecyl sulfate polyacryla mide gel electrophoresis separation. Polyclonal antibodies to these pr oteins were then produced in young male New Zealand White rabbits. bet a-Naphthoflavone (beta NF)- and phenobarbital (PB)-treated red-winged blackbird, screech owl, European starling, and lesser scaup liver micr osomes were analyzed in western blots for species cross-reactivity. Al though all four of these avian species exhibited cross-reactivity with antibodies to beta NF-induced mallard P450, all but the lesser scaup revealed a protein of higher molecular weight than that of the beta NF -induced mallard. In addition, only the lesser scaup exhibited cross-r eactivity with the anti-PB-induced mallard P450 antibodies.