Nalty T J, Yeoman L C
Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030.
J Immunol Methods. 1988 Feb 24;107(1):143-9. doi: 10.1016/0022-1759(88)90020-8.
A method is described for the silver staining of polyacrylamide gels and transfer to nitrocellulose sheets which couples the advantages of silver stain detection sensitivity with efficient transfer to nitrocellulose sheets. Using this procedure, a detection sensitivity of 10 ng was achieved while retaining a transfer efficiency for most proteins that is equivalent or better than methodology that does not permit prior visualization of protein. When coupled with the use of an acrylamide template, this procedure permits protein detection, simultaneous transfer of a large number of cored protein products from many one- or two-dimensional polyacrylamide gels to a single sheet of nitrocellulose, and retention of immunoreactivity. Prior detection, simultaneous transfer, and retention of immunoreactivity provide a substantial number of biochemical and technical advantages.
本文描述了一种用于聚丙烯酰胺凝胶银染及转印至硝酸纤维素膜的方法,该方法结合了银染检测灵敏度高和高效转印至硝酸纤维素膜的优点。使用此方法,检测灵敏度可达10 ng,同时对于大多数蛋白质而言,其转印效率与那些不允许蛋白质预先可视化的方法相当或更高。当与丙烯酰胺模板结合使用时,该方法可实现蛋白质检测、将大量来自多个一维或二维聚丙烯酰胺凝胶的核心蛋白质产物同时转印至单张硝酸纤维素膜,并保留免疫反应性。预先检测、同时转印和保留免疫反应性具有诸多生化和技术优势。