Marti U, Hauri H P, Meyer U A
Department of Pharmacology, University of Basel, Switzerland.
Eur J Cell Biol. 1990 Aug;52(2):193-200.
A panel of murine monoclonal antibodies was produced against phenobarbital-inducible cytochrome P450-PB of rat liver in order to establish specific immunological tools for studying the induction process in situ by immunoelectron microscopy and in vitro by a novel ELISA. Antibody 573/64 was found to be useful for both approaches. The immunolabeling procedure with protein A-colloidal gold applied to Lowicryl K4M-embedded rat liver revealed the rough ER as the primary site of cytochrome P450-PB induction. This organelle showed the highest labeling density 12 h after administration of phenobarbital while after maximal enzyme induction at day 5 the labeling density was highest in the smooth ER. Maximal increase in cytochrome P450-PB was 21-fold by morphometric analysis and 15-fold by ELISA. In addition, the enzyme apparently does neither recycle through the Golgi apparatus nor is it degraded in lysosomes when maximally induced.
制备了一组针对大鼠肝脏中苯巴比妥诱导型细胞色素P450 - PB的鼠单克隆抗体，以便建立特异性免疫工具，用于通过免疫电子显微镜原位研究诱导过程，并通过新型酶联免疫吸附测定（ELISA）进行体外研究。发现抗体573/64对这两种方法均有用。将蛋白A - 胶体金免疫标记程序应用于用Lowicryl K4M包埋的大鼠肝脏，结果显示粗面内质网是细胞色素P450 - PB诱导的主要部位。在给予苯巴比妥后12小时，该细胞器显示出最高的标记密度，而在第5天酶诱导达到最大值时，标记密度在滑面内质网中最高。通过形态计量分析，细胞色素P450 - PB的最大增加为21倍，通过ELISA为15倍。此外，当最大诱导时，该酶显然既不通过高尔基体循环，也不在溶酶体中降解。
