Chu Y H, Whitesides G M
Department of Chemistry, Harvard University, Cambridge, MA 02138.
Biotechniques. 1993 Jun;14(6):925-30.
Proteins stained with Coomassie Brilliant Blue I were transferred effectively by blotting from polyacrylamide gel electrophoresis (PAGE) gels to transparencies of the type used in plain-paper copiers. The details of the original electropherogram were retained on transfer and did not fade over a period of three years. Both the protein and the associated dye transfer; however, protein does not transfer in the absence of dye. Protein patterns present in several types of gels--sodium dodecyl sulfate (SDS)-PAGE, non-denaturing PAGE, isoelectric focusing PAGE and SDS-agarose--all transferred successfully after staining with dye 1. Proteins visualized with other organic dyes such as Fast Green FCF 2, Uniblue A 3 and Procion Blue MX-R 4 also transferred, but proteins stained with Stains-all 5 or silver did not. This transfer provides a simple, economical way to preserve data from slab gel electrophoresis and a convenient method to display data using an overhead projector. The blotted transparencies are also excellent substrates for use with gel scanning densitometers.
用考马斯亮蓝I染色的蛋白质通过印迹法能有效地从聚丙烯酰胺凝胶电泳(PAGE)凝胶转移至普通纸复印机所用类型的透明胶片上。原始电泳图谱的细节在转移后得以保留,并且在三年时间内都没有褪色。蛋白质和相关染料都能转移;然而,在没有染料的情况下蛋白质不会转移。存在于几种类型凝胶中的蛋白质模式——十二烷基硫酸钠(SDS)-PAGE、非变性PAGE、等电聚焦PAGE和SDS-琼脂糖——在用染料1染色后都成功转移。用其他有机染料如固绿FCF 2、优尼蓝A 3和普施安蓝MX-R 4可视化的蛋白质也能转移,但用全染剂5或银染色的蛋白质不能转移。这种转移提供了一种简单、经济的方法来保存平板凝胶电泳的数据,以及一种使用投影仪显示数据的便捷方法。印迹后的透明胶片也是与凝胶扫描密度计配合使用的优质底物。