组织培养分级分离。对经甘油诱导裂解匀浆的莱特雷细胞进行区带离心：在蔗糖梯度中对膜进行亚分级分离。
Tissue-culture fractionation. Zonal centrifugation of Lettrée cells homogenized by glycerol-induced lysis: subfractionation of membranes in sucrose gradients.
作者信息
Graham J M, Coffey K H
出版信息
Biochem J. 1979 Jul 15;182(1):165-71. doi: 10.1042/bj1820165.
Abstract
- Lettrée cells were grown intraperitoneally in MF-1 mice. 2. Cells that were loaded with glycerol were swollen in 0.1 M-sucrose and disrupted by Dounce homogenization. 3. Early-passage Lettrée cells were more easily disrupted than late-passage cells by this method, and the former produced larger fragments of plasma membrane. 4. The membranes were fractionated initially in sucrose gradients (on the basis of sedimentation rate) in a BXIV zonal rotor. 5. Fractions from this gradient were further resolved in isopycnic sucrose gradients. 6. Plasma-membrane and endoplasmic-reticulum fractions were recovered in good yield and high purity.
摘要
- 将Lettrée细胞接种于MF-1小鼠的腹腔内进行培养。2. 用甘油加载的细胞在0.1M蔗糖中肿胀，然后通过玻璃匀浆器匀浆破碎。3. 用这种方法，早期传代的Lettrée细胞比晚期传代的细胞更容易破碎，并且前者产生的质膜碎片更大。4. 首先在BXIV区带转子中通过蔗糖梯度（基于沉降速率）对膜进行分级分离。5. 从该梯度中收集的级分在等密度蔗糖梯度中进一步分离。6. 以高产量和高纯度回收质膜和内质网级分。
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