Manabe T, Miyamoto H, Oota H, Toyota Y
Department of Chemistry, Faculty of Science, Ehime University, Matsuyama, Japan.
Electrophoresis. 1998 Jun;19(8-9):1319-24. doi: 10.1002/elps.1150190819.
We used three different electrophoretic techniques for the analysis of human plasma proteins: (i) two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), with sodium dodecyl sulfate (SDS) used only in slab gel electrophoresis; (ii) capillary isoelectric focusing (CIEF) with no denaturants; (iii) linear polyacrylamide (LPA)-filled capillary electrophoresis with SDS (SDS-CE). With technique (i), data on isoelectric point and molecular size of plasma proteins can be obtained. Techniques (ii) and (iii) are suited to obtain quantitative information on proteins. The separation principle used in technique (ii) is closely related to that used in the first dimension of technique (i), and that used in technique (iii) related to that in the second dimension of technique (i). Therefore, we could successfully correlate protein separation patterns obtained by 2-D PAGE and those obtained by capillary electrophoresis. The advantages of correlating data obtained by various electrophoretic techniques in the course of constructing a comprehensive database on human plasma proteins are discussed.
(i)二维聚丙烯酰胺凝胶电泳(2-D PAGE),仅在平板凝胶电泳中使用十二烷基硫酸钠(SDS);(ii)无变性剂的毛细管等电聚焦(CIEF);(iii)含SDS的线性聚丙烯酰胺(LPA)填充毛细管电泳(SDS-CE)。使用技术(i)可获得血浆蛋白的等电点和分子大小数据。技术(ii)和(iii)适合获取蛋白质的定量信息。技术(ii)所采用的分离原理与技术(i)第一维所采用的原理密切相关,技术(iii)所采用的原理与技术(i)第二维所采用的原理相关。因此,我们能够成功地将2-D PAGE获得的蛋白质分离图谱与毛细管电泳获得的图谱相关联。本文讨论了在构建人血浆蛋白综合数据库过程中关联各种电泳技术所获数据的优势。