Shiba K S, Kanamori K, Cho H, Furuhata N, Harada T, Shiba A, Nakao M
Department of Clinical Biochemistry, Faculty of Medicine, Tokyo Medical and Dental University, Bunkyo-ku, Japan.
Clin Chim Acta. 1988 Feb 29;172(1):77-84. doi: 10.1016/0009-8981(88)90122-2.
Proteins in normal human urine were clearly fractionated into 26 bands with molecular weights from 14,000 to 230,000 by means of one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) coupled with silver staining. The main band contained uromucoid, and the second main band had albumin. However, when urine samples from healthy persons were electrophoresed in the absence of SDS using polyacrylamide gel or agarose gel, or a cellulose acetate membrane, albumin but not uromucoid, frequently formed the main protein band. It is suggested that this is due to the complexing of uromucoid subunits to form a large molecule which cannot penetrate into the gel. In order to correctly fractionate all the proteins contained in normal human urine, it was concluded that it was best to treat a urine sample with SDS with pre-condensation, fractionate it by SDS-PAGE and stain fractionated proteins by a highly sensitive method such as silver staining.
通过一维十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）结合银染法，正常人尿液中的蛋白质可清晰地分离成26条带，分子量范围为14,000至230,000。主带含有尿黏蛋白，第二条主带含有白蛋白。然而，当使用聚丙烯酰胺凝胶、琼脂糖凝胶或醋酸纤维素膜在无SDS的情况下对健康人的尿液样本进行电泳时，白蛋白而非尿黏蛋白常常形成主要蛋白带。这表明这是由于尿黏蛋白亚基复合形成了一个不能穿透凝胶的大分子。为了正确分离正常人尿液中所含的所有蛋白质，得出的结论是，最好用预浓缩的SDS处理尿液样本，通过SDS - PAGE进行分离，并用银染等高灵敏度方法对分离出的蛋白质进行染色。
