...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Analysis of proteins in biological samples by capillary sieving electrophoresis with postcolumn derivatization/laser-induced fluorescence detection.
【24h】

Analysis of proteins in biological samples by capillary sieving electrophoresis with postcolumn derivatization/laser-induced fluorescence detection.

机译：毛细管筛分电泳和柱后衍生/激光诱导荧光检测对生物样品中的蛋白质进行分析。

获取原文
获取原文并翻译 | 示例
   

获取外文期刊封面封底 >>

       
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Previously, we have demonstrated postcolumn derivatization of proteins separated by capillary sieving electrophoresis (CSE), in which naphthalene-2,3-dicarbaldehyde was employed as a fluorogenic labeling reagent. Standard proteins separated by CSE were reacted with naphthalene-2,3-dicarbaldehyde in the presence of 2-mercaptoethanol (2-ME) which plays a role of a reducing agent in the derivatization reaction. To improve the sensitivity, we attempted the use of ethanethiol instead of 2-ME. Ethanethiol showed 1.4- to 4.5-fold lower limits of detection for proteins than 2-ME. Furthermore, we found that 8-aminopyrene-1,3,6-trisulfonate (APTS) is a good marker for relative electrophoretic mobilities of proteins in CSE. Since APTS is a fluorescent and trivalent anion, it generates strong fluorescence and migrates faster than any of the proteins. Therefore, we employed APTS as a marker to obtain the relative electrophoretic mobilities of proteins. The present method was applied to the analyses of proteins in biological samples. Human Ewing's family tumor cell line 'RDES' was used as a sample. The cultured cells were lysed with a buffer containing Tris-HCl, NaCl, sodium dodecyl sulfate, and 2-ME. After denaturation, the lysate was directly introduced into the capillary. Several peaks, which would correspond to proteins with molecular mass ranging from 10 to 93 kDa, were found in the cell lysate. In addition, we measured a milk sample by the CSE with postcolumn derivatization. The electropherogram showed five major peaks which corresponded to alpha-lactalbumin, beta-lactoglobulin, kappa-casein, bovine serum albumin, and mixture of alpha- and beta-casein.
机译：以前，我们已经证明了通过毛细管筛分电泳（CSE）分离的蛋白质的柱后衍生化，其中使用萘2,3-二甲醛作为荧光标记试剂。在2-巯基乙醇（2-ME）的存在下，通过CSE分离的标准蛋白质与萘-2,3-二甲醛反应，该衍生物在衍生化反应中起还原剂的作用。为了提高灵敏度，我们尝试使用乙硫醇代替2-ME。乙硫醇的蛋白质检出限比2-ME低1.4-4.5倍。此外，我们发现8-氨基py-1,3,6-三磺酸盐（APTS）是CSE中蛋白质相对电泳迁移率的良好标记。由于APTS是一种荧光和三价阴离子，因此它会产生强荧光，并且比任何一种蛋白质迁移都快。因此，我们采用APTS作为标记来获得蛋白质的相对电泳迁移率。本方法应用于生物样品中蛋白质的分析。人类尤因氏家族肿瘤细胞系“ RDES”被用作样品。用含有Tris-HCl，NaCl，十二烷基硫酸钠和2-ME的缓冲液裂解培养的细胞。变性后，将裂解物直接引入毛细管。在细胞裂解物中发现了几个峰，这些峰对应于分子量范围为10至93 kDa的蛋白质。此外，我们通过CSE在柱后衍生化中测量了牛奶样品。电泳图显示五个主要峰，分别对应于α-乳白蛋白，β-乳球蛋白，κ-酪蛋白，牛血清白蛋白以及α-酪蛋白和β-酪蛋白的混合物。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号