Cubillos-Rojas Monica, Amair-Pinedo Fabiola, Tato Irantzu, Bartrons Ramon, Ventura Francesc, Rosa Jose Luis
Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain.
Methods Mol Biol. 2012;869:205-13. doi: 10.1007/978-1-61779-821-4_17.
Polyacrylamide gel electrophoresis (PAGE) is one of the most powerful tools used for protein analysis. We describe the use of Tris-acetate buffer and 3-15% polyacrylamide gradient gels to simultaneously separate proteins in the mass range of 10-500 kDa. We show that this system is highly sensitive, it has good resolution and high reproducibility, and that it can be used for general applications of PAGE such as Coomassie Brilliant Blue staining and immunoblotting. Moreover, we describe how to generate mini Tris-acetate polyacrylamide gels to use them in miniprotein electrophoresis systems. These economical gels are easy to generate and to manipulate and allow a rapid analysis of proteins. All these features make the Tris-acetate-PAGE system a very helpful tool for protein analysis.
聚丙烯酰胺凝胶电泳(PAGE)是用于蛋白质分析的最强大工具之一。我们描述了使用Tris-乙酸缓冲液和3-15%聚丙烯酰胺梯度凝胶同时分离质量范围为10-500 kDa的蛋白质。我们表明,该系统高度灵敏,具有良好的分辨率和高重现性,可用于PAGE的常规应用,如考马斯亮蓝染色和免疫印迹。此外,我们还描述了如何制备小型Tris-乙酸聚丙烯酰胺凝胶以用于小型蛋白质电泳系统。这些经济实惠的凝胶易于制备和操作,可快速分析蛋白质。所有这些特性使Tris-乙酸-PAGE系统成为蛋白质分析的非常有用的工具。