RAPID ISOLATION OF PLASMA-MEMBRANES IN HIGH-YIELD FROM CULTURED FIBROBLASTS
RAPID ISOLATION OF PLASMA-MEMBRANES IN HIGH-YIELD FROM CULTURED FIBROBLASTS
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DOI：
10.1042/bj1680187
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发表时间：
1977-01-01
影响因子：
4.1
通讯作者：
REES, DA
中科院分区：
文献类型：
--
作者：
THOM, D;POWELL, AJ;REES, DA
开发了一种方法，该方法允许从培养的[大鼠皮肤]成纤维细胞中以高产率快速制备纯质膜。在高渗硼酸盐/EDTA中裂解细胞，差速离心后，在蔗糖屏障上离心收集膜。分离材料的EM显示基本上不含污染细胞器的大膜囊泡。没有可检测到的内质网酶标记物NADH 2-硫辛酰胺氧化还原酶（EC 1.6.4.3）的活性，并且线粒体标记物琥珀酸脱氢酶（EC 1.3.99.1）的活性显著降低。化学成分与以前的观察结果吻合得很好。应用同位素标记分离质膜的有用性得到了证实。
A method was developed which allows the rapid preparation of pure plasma membranes in high yield from cultured [rat dermal] fibroblasts. Cells are lysed in hypt-osmotic borate/EDTA and, after differential centrifugation, the membranes collected by centrifugation on a sucrose barrier. EM of the isolated material shows large membrane vesicles essentially free from contaminating organelles. There is no detectable activity of the endoplasmic-reticulum enzyme marker, NADH2-lipoamide oxidoreductase (EC 1.6.4.3), and that of succinate dehydrogenase (EC 1.3.99.1), a marker for mitochondria, is substantially decreased. Chemical compositions are in good agreement with previous observations. The usefulness of applied isotopic markers for isolating plasma membranes was confirmed.