Kyhse-Andersen J
J Biochem Biophys Methods. 1984 Dec;10(3-4):203-9. doi: 10.1016/0165-022x(84)90040-x.
A simple, horizontal device for rapid electrophoretic transfer of proteins from several polyacrylamide gels simultaneously is described. Up to six 'TRANS-UNITS' consisting of soaked filter paper on either side of polyacrylamide gel/nitrocellulose sheets that are separated by dialysis membranes are stacked between graphite plate electrodes. The only buffer reservoir in the apparatus is that in stacked, soaked filter paper. A special buffer system based on the isotachophoresis theory was developed for this purpose. The electrophoretic transfer was performed with equal efficiency in all TRANS-UNITS of the stack. Only traces of a few proteins remained in the polyacrylamide gel after transfer. With this apparatus, 50 protein bands from a human serum protein sample (diluted 1 : 100) were detected by immunoblotting with the retainment of the high resolution of the SDS-PAGE technique. The apparatus provided a constant current density of 0.8 mA/cm2 during the 1-h transfer time at 21 degrees C, irrespective of the number of TRANS-UNITS. The apparatus generated 1-5 W in joule heat, depending on the number of TRANS-UNITS in the stack.
本文描述了一种简单的水平装置，可同时将蛋白质从多个聚丙烯酰胺凝胶中快速进行电泳转移。由透析膜隔开的聚丙烯酰胺凝胶/硝酸纤维素片两侧的浸湿滤纸组成的多达六个“转移单元”堆叠在石墨板电极之间。该装置中唯一的缓冲液储存器是堆叠的浸湿滤纸中的缓冲液。为此开发了一种基于等速电泳理论的特殊缓冲系统。在堆叠的所有转移单元中，电泳转移均以相同效率进行。转移后，聚丙烯酰胺凝胶中仅残留少量蛋白质痕迹。使用该装置，通过免疫印迹法检测了来自人血清蛋白样品（稀释1:100）的50条蛋白带，同时保留了SDS-PAGE技术的高分辨率。在21摄氏度下1小时的转移时间内，该装置提供0.8 mA/cm2的恒定电流密度，与转移单元的数量无关。根据堆叠中转移单元的数量，该装置产生1 - 5瓦的焦耳热。
