Braun W, Abraham R
Department of Medical Virology, University Clinics of Frankfurt.
Electrophoresis. 1989 Apr;10(4):249-53. doi: 10.1002/elps.1150100406.
An easy-to-perform and efficient blotting method was developed for the transfer of proteins separated in PhastGel media. The method was evaluated by comparing the blotting results of low molecular weight marker proteins at different times. 14C-Labelled proteins were used for the assessment of the transfer efficiency. Highly efficient transfer was achieved within 1 h of blotting and the proteins were almost completely eluted from the gel in 2 h. The gel remained on its solid plastic backing, which is not the case when using an electrophoretic transfer method. As little as 1.25 ng of electrophoretically separated protein were enzymatically detected on the blot when applying blot-biotinylation. The transfer was performed at low temperature (4 degrees C), which-in contrast to thermoblotting-additionally offers the chance to blot thermolabile proteins. These superior features, together with the method's versatility and low cost, mark an interesting alternative to the previously recommended blotting procedures after PhastSystem electrophoresis.
我们开发了一种易于操作且高效的印迹方法，用于转移在PhastGel介质中分离的蛋白质。通过比较不同时间低分子量标记蛋白的印迹结果对该方法进行了评估。使用14C标记的蛋白质评估转移效率。在印迹1小时内实现了高效转移，并且蛋白质在2小时内几乎完全从凝胶中洗脱。凝胶仍保留在其固体塑料背衬上，而使用电泳转移方法时则不然。应用印迹生物素化时，在印迹上酶法检测到低至1.25 ng的电泳分离蛋白。转移在低温（4摄氏度）下进行，与热印迹相比，这还为印迹热不稳定蛋白提供了机会。这些优越的特性，再加上该方法的多功能性和低成本，使其成为PhastSystem电泳后先前推荐的印迹程序的一个有趣替代方案。
