Preparative electrophoresis of living mammalian cells in a stationary ficoll gradient.
Preparative electrophoresis of living mammalian cells in a stationary ficoll gradient.
复制标题
在固定聚蔗糖梯度中对活哺乳动物细胞进行制备型电泳。
DOI：
10.1080/00327487308061523
复制
发表时间：
1973
期刊：
影响因子：
--
通讯作者：
M. K. Louie
中科院分区：
文献类型：
--
作者：
R. Boltz;P. Todd;M. Streibel;M. K. Louie
摘要设计了一种用于哺乳动物活细胞垂直密度梯度电泳的装置。培养的中国仓鼠细胞（M3- 1F 3）在悬浮液中生长，并在0-10% ficoll梯度上分层，该梯度也是磷酸盐缓冲液pB 7.2和1 Z葡萄糖中的6.8-5.1%蔗糖梯度。该组合物的60-ml垂直梯度在2.3 cm直径的玻璃柱中在15 cm的距离上覆盖1.04-1.05 g/cm 3的密度范围。施加约2.3V/cm的电场5小时，在此期间细胞迁移4.7cm。迁移率与电泳迁移率为-1.15 ± 0.20 μm/sec/volt/cm的细胞一致，与微电泳测定的迁移率相等。在电泳后，以50个级分收获梯度，并且基于集落形成，细胞的存活率为75%。
Abstract An apparatus was designed for the vertical density-gradient electrophoresis of viable mammalian cells. Cultured Chinese hamster cells (M3-1F3) were grown is suspension and layered on top of a 0–10% ficoll gradient which was also an inverse 6.8-5.1% sucrose gradient in phosphate buffer, pB 7.2 and 1Z glucose. A 60-ml vertical gradient of this composition covered the density range 1.04–1.05 g/cm3 over a distance of 15 cm in a 2.3 cm diameter glass column. An electric field of approximately 2.3 V/cm was applied for 5 hr. The cells migrated 4.7 cm during this period. The migration rate was consistent with the cells having an electrophoretic mobility of -1.15 ± 0.20 μm/sec per volt/cm, equal to that determined by microelectrophoresis. The gradient was harvested in 50 fractions after electrophoresis, and the viability of the cells was 75% on the basis of colony formation.