Hall Randy A
Department of Pharmacology, Emory University School of Medicine, Rollins Research Center, Room 5113, 1510 Clifton Road, Atlanta, 30322, GA, USA,
Methods Mol Biol. 2015;1278:371-9. doi: 10.1007/978-1-4939-2425-7_24.
Blot overlay is a useful method for studying protein-protein interactions. This technique involves fractionating proteins on SDS-PAGE, blotting to nitrocellulose or PVDF membrane, and then incubating with a probe of interest. The probe is typically a protein that is radiolabeled, biotinylated, or simply visualized with a specific antibody. When the probe is visualized via antibody detection, this technique is often referred to as "Far Western blot." Many different kinds of protein-protein interactions can be studied via blot overlay, and the method is applicable to screens for unknown protein-protein interactions as well as to the detailed characterization of known interactions.
印迹覆盖法是研究蛋白质 - 蛋白质相互作用的一种有用方法。该技术包括在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)上分离蛋白质,将其印迹到硝酸纤维素膜或聚偏二氟乙烯(PVDF)膜上,然后与感兴趣的探针孵育。探针通常是一种经过放射性标记、生物素化或简单地用特异性抗体可视化的蛋白质。当通过抗体检测使探针可视化时,这种技术通常被称为“Far Western印迹法”。许多不同类型的蛋白质 - 蛋白质相互作用都可以通过印迹覆盖法进行研究,该方法适用于未知蛋白质 - 蛋白质相互作用的筛选以及已知相互作用的详细表征。