Galteau M M, Antoine B, Reggio H
EMBO J. 1985 Nov;4(11):2793-800. doi: 10.1002/j.1460-2075.1985.tb04005.x.
Epoxide hydrolase (EH, EC 3.3.2.3) was chosen as a potential marker for smooth endoplasmic reticulum, because this enzyme is inducible by drugs such as phenobarbital. The hypothesis was verified in rat liver using immunochemical and immunocytochemical techniques. Antibodies were raised to the purified protein. These antibodies were affinity purified using the enzyme immobilized on Sepharose Ultrogel. The specificity of the antibodies was assayed by immunoelectrotransfer (Western blot). The labelling of rat liver thin frozen sections with protein A-gold particles demonstrated that the antibodies specifically recognised smooth endoplasmic reticulum membranes. Rough endoplasmic reticulum, other intracellular organelles and plasma membrane were unlabelled.
环氧水解酶(EH,EC 3.3.2.3)被选为滑面内质网的潜在标志物,因为该酶可被苯巴比妥等药物诱导。使用免疫化学和免疫细胞化学技术在大鼠肝脏中验证了这一假设。制备了针对纯化蛋白的抗体。这些抗体使用固定在琼脂糖凝胶上的酶进行亲和纯化。通过免疫电转移(蛋白质印迹法)检测抗体的特异性。用蛋白A-金颗粒标记大鼠肝脏薄冰冻切片表明,抗体特异性识别滑面内质网膜。粗面内质网、其他细胞内细胞器和质膜未被标记。