使用一维和二维毛细管电泳分析AtT - 20细胞匀浆的不同去污剂组分。

Analysis of differential detergent fractions of an AtT-20 cellular homogenate using one- and two-dimensional capillary electrophoresis.

作者信息

Fazal Md Abul, Palmer Vanessa R, Dovichi Norman J

机构信息

Department of Chemistry, University of Washington, Seattle, 98195-1700, USA.

出版信息

J Chromatogr A. 2006 Oct 20;1130(2):182-9. doi: 10.1016/j.chroma.2006.05.053. Epub 2006 Jun 16.

DOI:10.1016/j.chroma.2006.05.053

PMID:16781720

Abstract

Differential detergent fractionation was used to sequentially extract cytosolic, membrane, nuclear, and cytoskeletal fractions from AtT-20 cells. Extracted components were denatured by sodium dodecyl sulfate (SDS) and then labeled with the fluorogenic reagent 3-(2-furoyl) quinoline-1-carboxaldehyde. Both capillary sieving electrophoresis (CSE) and micellar electrokinetic capillary chromatography (MECC) were used to separate labeled components by one-dimensional (1D) electrophoresis. Labeled components were also separated by two-dimensional (2D) capillary electrophoresis; CSE was employed in the first dimension and MECC in the second dimension. Roughly 150 fractions were transferred from the first to the second capillary for this comprehensive analysis in 2.5 h.

摘要

采用差异去污剂分级分离法从AtT-20细胞中依次提取胞质、膜、核和细胞骨架组分。提取的组分用十二烷基硫酸钠（SDS）变性，然后用荧光试剂3-（2-呋喃甲酰基）喹啉-1-甲醛标记。毛细管筛分电泳（CSE）和胶束电动毛细管色谱法（MECC）均用于通过一维（1D）电泳分离标记的组分。标记的组分也通过二维（2D）毛细管电泳分离；第一维采用CSE，第二维采用MECC。在2.5小时内，大约150个级分从第一根毛细管转移到第二根毛细管进行这种全面分析。