Wirth P J, Romano A
Biopolymer Chemistry Section, Laboratory of Experimental Carcinogenesis, National Cancer Institute, Bethesda, MD 20892, USA.
J Chromatogr A. 1995 Apr 28;698(1-2):123-43. doi: 10.1016/0021-9673(94)00879-e.
Polyacrylamide gel electrophoresis is a reliable and widely used technique for the separation, identification and characterization of proteins and protein mixtures. With the introduction of high resolution two-dimensional polyacrylamide gel electrophoresis in 1975 upward to 2000 individual polypeptides spots are easily separated on a single electrophoretic gel thereby necessitating the availability of highly sensitive protein detection methods. Although a plethora of protein-staining and -visualization protocols have been described utilizing both radioactive and non-radioactive reagents, many times the use of mono-dimensional detection procedures is insufficient to address the experimental questions asked. The present review highlights the utilization of combined protein-labeling and -staining methodologies in gel electrophoresis including selected applications in polyacrylamide gels and solid membrane matrixes.
聚丙烯酰胺凝胶电泳是一种可靠且广泛应用的技术，用于蛋白质及蛋白质混合物的分离、鉴定和表征。自1975年引入高分辨率二维聚丙烯酰胺凝胶电泳以来，在一块电泳凝胶上可轻松分离多达2000个单个多肽斑点，因此需要有高灵敏度的蛋白质检测方法。尽管已经描述了大量使用放射性和非放射性试剂的蛋白质染色和可视化方案，但很多时候，一维检测程序不足以解决所提出的实验问题。本综述重点介绍了凝胶电泳中蛋白质标记和染色相结合的方法的应用，包括在聚丙烯酰胺凝胶和固体膜基质中的特定应用。
