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标题：DELICATE AND CRUCIAL PH DEPENDENCE IN PERMEABILIZATION OF CULTURED MAMMALIAN CELLS BY VORTEX-STIRRING WITH A HIGH MOLECULAR WEIGHT POLYACRYLIC ACID
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作者：Naoki SHIMIZU ; Yutaka KAWAZOE
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：1996
卷号：19
期号：11
页码：1533-1535
DOI：10.1248/bpb.19.1533
出版社：The Pharmaceutical Society of Japan
摘要：We previously reported that cultured mammalian cells are effectively permeabilized by vortex-stirring the cells with a high molecular weight polyacrylic acid. The present study revealed that the efficiency of permeabilization of a non-permeant dye, Lucifer Yellow (LY), was sensitively affected by the pH of the medium. When the cells are vortex-stirred in RPMI 1640 culture medium, the pH of the medium should be adjusted with HEPES and NaOH, in stead of NaHCO₃, to a pH higher than 7.6 for successful permeabilization; the higher the pH, the better the result obtained. Internalization of LY was near the background level at a pH below 7.4. When phosphate-buffered saline was substituted for RPMI 1640 medium, the optimal pH range was slightly shifted to a more acidic region. This requirement for the pH of the medium is indispensable as a supplement to the standard permeabilization procedure tentatively proposed in the preceding report : Biol. Pharm. Bull., 19, 1279-1284 (1996).
关键词：permeabilization;polycrylic acid;pH of medium;Lucifer Yellow;cultured cell