Klafki H W, Wiltfang J, Staufenbiel M
Preclinical Research, Sandoz Pharma Ltd., Basel, CH-4002, Switzerland.
Anal Biochem. 1996 May 15;237(1):24-9. doi: 10.1006/abio.1996.0195.
Different sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) systems designed for the separation of peptides were compared for their usefulness in separating synthetic beta-amyloid peptides betaA4 (1-40) and betaA4 (1-42). Clear resolution was achieved by addition of 8 M urea to the separation gel and use of a multiphasic buffer system employing bicine and sulfate as trailing and leading ions, respectively (bicine/Tris/urea gels). Under these conditions, the longer peptide migrated faster than the one ending at amino acid 40. The usefulness of this SDS-PAGE system for the analysis of betaA4-related peptides generated during cellular metabolism was demonstrated by immunoprecipitation and electrophoretic separation of radiolabeled peptides secreted by cells transfected with amyloid precursor protein cDNAs.
比较了为分离肽段而设计的不同十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）系统在分离合成β-淀粉样肽βA4（1-40）和βA4（1-42）方面的实用性。通过在分离胶中添加8M尿素，并使用分别以二乙醇胺和硫酸盐作为尾随离子和前导离子的多相缓冲系统（二乙醇胺/Tris/尿素凝胶），实现了清晰的分离效果。在这些条件下，较长的肽段比较短的（在氨基酸40处终止的肽段）迁移速度更快。通过对用淀粉样前体蛋白cDNA转染的细胞分泌的放射性标记肽段进行免疫沉淀和电泳分离，证明了该SDS-PAGE系统在分析细胞代谢过程中产生的βA4相关肽段方面的实用性。
