Nguyen T D, Broyart J P, Ngu K T, Illescas A, Mircheff A K, Gray G M
Department of Medicine, Stanford University, California 94305.
J Membr Biol. 1987;98(3):197-205. doi: 10.1007/BF01871183.
A simplified method for isolating highly purified laterobasal membranes (LBM) from enterocytes is based on treatment of membranes with 8 mM CaCl2 concentration in order to aggregate intracellular membrane contaminants. The resultant LBM showed an average 15-fold enrichment and constituted 8% of the original K-stimulated phosphatase in the initial crude homogenate. It showed typical LBM migration on counter-current distribution (CCD) and was essentially free of contamination with endoplasmic reticulum and Golgi membranes. This method is highly efficient and yields sufficient purified LBM to allow comprehensive analysis of enterocyte membrane events.
一种从肠上皮细胞中分离高纯度侧基底膜（LBM）的简化方法是基于用8 mM氯化钙浓度处理膜，以聚集细胞内膜污染物。所得的LBM显示平均富集了15倍，占初始粗匀浆中原始K刺激磷酸酶的8%。它在逆流分配（CCD）中显示出典型的LBM迁移，并且基本没有内质网和高尔基体膜的污染。该方法效率很高，可产生足够的纯化LBM，以便对肠上皮细胞膜事件进行全面分析。
