Biber J, Rechkemmer G, Bodmer M, Schröder P, Haase W, Murer H
Biochim Biophys Acta. 1983 Oct 26;735(1):1-11. doi: 10.1016/0005-2736(83)90255-9.
A method for an analytical isolation of plasma membranes from columnar cells (colonocytes) of the proximal colon of the guinea pig is described. Isolation of the colonocytes was performed by a mild EDTA-chelation method. After homogenization, two subsequent sucrose gradient centrifugations (isokinetic and isopycnic) yielded a plasma-membrane fraction which was enriched 12-fold in (Na+ + K+)-ATPase activity and 8-fold in adenylate cyclase activity. It is suggested that the purified membrane fraction consists mainly of basolateral membranes of the colonocytes. Due to the lack of suitable marker enzymes, no evidence for enrichment of the brush-border membranes was obtained. Histochemical studies demonstrated that alkaline phosphatase is absent from the luminal membrane of the surface epithelial cells of the proximal colon of the guinea pig.
本文描述了一种从豚鼠近端结肠柱状细胞（结肠上皮细胞）中分析性分离质膜的方法。采用温和的乙二胺四乙酸（EDTA）螯合方法分离结肠上皮细胞。匀浆后，通过两次连续的蔗糖梯度离心（等动力学和等密度）得到一个质膜组分，其（钠+钾）-ATP酶活性富集了12倍，腺苷酸环化酶活性富集了8倍。提示纯化的膜组分主要由结肠上皮细胞的基底外侧膜组成。由于缺乏合适的标记酶，未获得刷状缘膜富集的证据。组织化学研究表明，豚鼠近端结肠表面上皮细胞腔面膜不存在碱性磷酸酶。
