Iwata N, Mukai T, Hara S, Endo T, Tomoda A
Department of Forensic Medicine, Tokyo Medical College.
Tohoku J Exp Med. 1996 Sep;180(1):65-71. doi: 10.1620/tjem.180.65.
We could prepare the microsomal fraction of mouse liver, without using an ultracentrifuge but with a low speed centrifuge. The procedure includes 1) lyophilization of post-mitochondrial fraction (9,000 x g supernatant) of mouse liver, 2) powdering of the lyophilized sample, 3) the addition of 1.15 per cent potassium chloride solution or distilled water, which afforded microsomal aggregates, 4) sedimentation of microsomal fraction by low-speed centrifugation (20,000 x g, 20 min). The sedimented microsomal fraction showed normal contents of cytochrome P-450 and cytochrome b5, and gave a normal pattern on SDS polyacrylamide gel electrophoresis and normal electron microscopic feature. This method should be convenient for rapid and large-scale preparation of microsomes, especially for the preparation of cytochrome b5 and cytochrome P-450.
我们可以不使用超速离心机，而是用低速离心机来制备小鼠肝脏的微粒体部分。该步骤包括：1）冻干小鼠肝脏的线粒体后部分（9,000×g上清液）；2）将冻干后的样品研磨成粉末；3）加入1.15%的氯化钾溶液或蒸馏水，从而形成微粒体聚集体；4）通过低速离心（20,000×g，20分钟）沉淀微粒体部分。沉淀的微粒体部分显示出细胞色素P-450和细胞色素b5的正常含量，在SDS聚丙烯酰胺凝胶电泳上呈现正常图谱，并且具有正常的电子显微镜特征。该方法对于快速大规模制备微粒体应该很方便，特别是用于制备细胞色素b5和细胞色素P-450。
