Lee C, Levin A, Branton D
Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
Anal Biochem. 1987 Nov 1;166(2):308-12. doi: 10.1016/0003-2697(87)90579-3.
We present a new method for visualizing proteins electrophoresed in sodium dodecyl sulfate-polyacrylamide gels. After electrophoresis, gels are incubated in CuCl2 to produce a negative image of colorless protein bands against a semiopaque background. Gels are stained completely within 5 min, do not require destaining, and can be stored indefinitely without loss of the image. Because proteins are not permanently fixed within the gel, they can be quantitatively eluted after chelation of Cu with EDTA. The sensitivity of the CuCl2 stain falls between that of Coomassie blue and silver. We anticipate that CuCl2 will be useful in the rapid analysis of proteins by polyacrylamide gel electrophoresis and in the preparation of purified polypeptides by elution from gel slices.
我们提出了一种可视化十二烷基硫酸钠-聚丙烯酰胺凝胶中电泳蛋白质的新方法。电泳后,将凝胶在氯化铜中孵育,以在半透明背景上产生无色蛋白条带的负像。凝胶在5分钟内完全染色,无需脱色,并且可以无限期保存而不会损失图像。由于蛋白质不会永久固定在凝胶中,因此在铜与乙二胺四乙酸螯合后可以定量洗脱。氯化铜染色的灵敏度介于考马斯亮蓝和银之间。我们预计氯化铜将有助于通过聚丙烯酰胺凝胶电泳快速分析蛋白质,以及通过从凝胶切片中洗脱来制备纯化的多肽。