Sasse Joachim, Gallagher Sean R
Shriners Hospital for Crippled Children, Tampa, Florida.
UVP, Inc, Upland, California.
Curr Protoc Immunol. 2008 Nov;Chapter 8:8.10B.1-8.10B.6. doi: 10.1002/0471142735.im0810bs83.
Staining of blot transfer membranes permits visualization of proteins and allows the extent of transfer to be monitored. In the protocols described in this unit, proteins are stained after electroblotting from one-dimensional or two-dimensional polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. Protocols are provided for the use of six general protein stains: Amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, and India ink. In addition, the fluorescent stains fluorescamine and IAEDANS, which covalently react with bound proteins, are described. Approximate detection limits for each nonfluorescent stain are indicated along with membrane compatibilities.
印迹转移膜的染色可使蛋白质可视化，并能监测转移的程度。在本单元所述的实验方案中，蛋白质从一维或二维聚丙烯酰胺凝胶电转印至印迹膜（如聚偏二氟乙烯（PVDF）、硝酸纤维素或尼龙膜）后进行染色。提供了六种常用蛋白质染色剂的使用方案：氨基黑、考马斯亮蓝、丽春红S、胶体金、胶体银和印度墨汁。此外，还介绍了与结合蛋白发生共价反应的荧光染色剂荧光胺和碘乙酰胺基丹磺酰氯。文中指出了每种非荧光染色剂的大致检测限以及与膜的兼容性。
