Enhancement of 3-methylcholanthrene-mediated induction of cytochrome P-450 by 17 alpha-ethynylestradiol: immunochemical studies.
Enhancement of 3-methylcholanthrene-mediated induction of cytochrome P-450 by 17 alpha-ethynylestradiol: immunochemical studies.
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17 α-乙炔雌二醇增强 3-甲基胆蒽介导的细胞色素 P-450 诱导：免疫化学研究。
DOI：
10.1016/0006-2952(89)90079-8
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发表时间：
1989
影响因子：
5.8
通讯作者：
Sinclair,P
中科院分区：
文献类型：
--
作者：
Sundstrom,S;Sinclair,J;Smith,EL;Sinclair,P
Materials and methodsPrimary cultures of livers from 16-to 17-day-old chicken embryos were prepared and treated with MC (0.93 pM) and EEs (15 PM) as described previously [4]. Microsomes were prepared from cultured cells, and cytochrome P-450 and EROD were measured in the microsomes as described previously [4]. Protein was determined by the method of Lowry et al.[6], using bovine serum albumin as a standard. Microsomal proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in a 5-10% gradient gel, and immunoblots were prepared as described [5]. Immunoinhibition of EROD activity was determined by the method of Lubet et al.[7]. Briefly, liver microsomes from cultured chick embryo hepatocytes (25 ng protein/reaction) were incubated with increasing amounts of preimmune serum or serum from rabbits immunized with either purified cytochrome P-450r., cs7x, or with a 50K cytochrome P-450 purified from glutethimide-treated embryos (P-4500LuTSOK). Preimmune serum was present in each mixture to maintain a constant amount of serum. After 20 min of incubation at room temperature, EROD activity was measured as described previously [4].
