Kshirsagar B, Wiggins R C
Clin Chim Acta. 1986 Jul 15;158(1):13-22. doi: 10.1016/0009-8981(86)90111-7.
A sensitive one-dimensional SDS-polyacrylamide gel electrophoretic system was devised whereby the proteins in 1 microliter of unconcentrated urine could be visualized by silver staining over the range 9,000-900,000 molecular weight. Identification of urine proteins was confirmed by Western blotting using peroxidase labelled antibodies. A map of the major proteins visualized in urine from individuals with renal disease was constructed. We conclude that the information available from the simple analysis of proteins according to their size is limited to general conclusions regarding whether proteinuria is likely to be of tubular or glomerular or mixed origin. More specific identification of individual proteins is not feasible because simple protein staining is not sufficiently reliable to identify individual proteins. The reasons for this conclusion are as follows: many proteins in urine migrate with similar apparent molecular weights, some proteins are not visualized by silver staining, and albumin polymeric complexes and fragments can be present at almost any molecular weight. However, one-dimensional SDS-polyacrylamide gel electrophoresis together with Western blotting does provide reliable information which might be clinically and experimentally useful.
设计了一种灵敏的一维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳系统，通过该系统，1微升未浓缩尿液中的蛋白质在9000至900000分子量范围内可通过银染法显现出来。使用过氧化物酶标记抗体的蛋白质印迹法证实了尿液蛋白质的鉴定。构建了患有肾脏疾病个体尿液中可见主要蛋白质的图谱。我们得出结论，根据蛋白质大小进行简单分析所获得的信息仅限于关于蛋白尿可能是肾小管性、肾小球性还是混合性来源的一般性结论。由于简单的蛋白质染色不足以可靠地鉴定单个蛋白质，因此对单个蛋白质进行更具体的鉴定是不可行的。得出该结论的原因如下：尿液中的许多蛋白质以相似的表观分子量迁移，一些蛋白质不能通过银染显现出来，并且白蛋白聚合物复合物和片段几乎可以出现在任何分子量处。然而，一维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳与蛋白质印迹法相结合确实提供了可能在临床和实验中有用的可靠信息。
