组织培养细胞分级分离。通过甘油诱导裂解使组织培养细胞匀浆后对膜进行分级分离。
Tissue-culture cell fractionation. Fractionation of membranes from tissue-culture cells homogenized by glycerol-induced lysis.
作者信息
Graham J M, Sandall J K
出版信息
Biochem J. 1979 Jul 15;182(1):157-64. doi: 10.1042/bj1820157.
Abstract
- The disruption of various types of tissue-culture cells by (a) incubation in solutions of 1.2 M-glycerol and (b) transfer of the glycerol-loaded cells to relatively hypo-osmotic solutions of 0.25 M-sucrose was studied. 2. Bivalent cations (2mM-Mg2+) were generally included to preserve the nuclei, but some cells (polyoma-virus-transformed baby-hamster kidney cells) failed to be disrupted adequately under these conditions. 3. Other cells (mouse-embryo fibroblasts) required additional gentle Dounce homogenization to effect complete cell breakage. 4. Purification of the whole homogenate was carried out by a combination of differential centrifugation and sedimentation or flotation through sucrose gradients. 5. Enzyme analysis showed that plasma-membrane, endoplasmic-reticulum and mitochondrial fractions were obtained in good yield and purity.
摘要
- 研究了通过以下方式对各种类型的组织培养细胞进行裂解：(a) 在1.2 M甘油溶液中孵育，以及(b) 将加载了甘油的细胞转移至0.25 M蔗糖的相对低渗溶液中。2. 通常会加入二价阳离子（2 mM Mg2+）以保存细胞核，但某些细胞（多瘤病毒转化的幼仓鼠肾细胞）在这些条件下未能被充分裂解。3. 其他细胞（小鼠胚胎成纤维细胞）需要额外进行轻柔的杜恩斯匀浆以实现完全的细胞破碎。4. 通过差速离心以及通过蔗糖梯度进行沉降或浮选相结合的方法对整个匀浆进行纯化。5. 酶分析表明，获得了高产率和高纯度的质膜、内质网和线粒体组分。
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本文引用的文献
1
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2
SEPARATION AND PARTIAL PURIFICATION OF PLASMA-MEMBRANE FRAGMENTS FROM EHRLICH ASCITES CARCINOMA MICROSOMES.从艾氏腹水癌微粒体中分离和部分纯化质膜片段
Science. 1965 Jun 4;148(3675):1343-5. doi: 10.1126/science.148.3675.1343.
3
Microsomal triphosphopyridine nucleotide-cytochrome c reductase of liver.肝脏微粒体三磷酸吡啶核苷酸-细胞色素c还原酶
J Biol Chem. 1962 Feb;237:587-95.
4
The isolation of a cell membrane fraction from rat liver.从大鼠肝脏中分离出细胞膜组分。
J Biophys Biochem Cytol. 1960 Oct;8(2):413-22. doi: 10.1083/jcb.8.2.413.
5
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Virology. 1959 Jul;8(3):396-7. doi: 10.1016/0042-6822(59)90043-1.
6
A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid.用于比色法测定脱氧核糖核酸的二苯胺反应的条件及机制研究。
Biochem J. 1956 Feb;62(2):315-23. doi: 10.1042/bj0620315.
7
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J Biol Chem. 1970 Dec 10;245(23):6357-65.
8
The determination of nucleic acids.核酸的测定
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