Hall Randy A
Department of Pharmacology, Emory University, Atlanta, GA, USA.
Methods Mol Biol. 2004;261:167-74. doi: 10.1385/1-59259-762-9:167.
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SDS - PAGE上分离蛋白质,印迹到硝酸纤维素膜或PVDF膜上,然后与感兴趣的探针孵育。探针通常是放射性标记、生物素化的蛋白质,或简单地用特异性抗体进行可视化检测。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”。通过印迹覆盖法可以研究许多不同类型的蛋白质 - 蛋白质相互作用,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。