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Capillary sodium dodecyl sulfate‐DALT electrophoresis with laser‐induced fluorescence detection for size‐based analysis of proteins in human colon cancer cells
Author(s) -
Hu Shen,
Jiang Jiang,
Cook Lillian M.,
Richards Dawn P.,
Horlick Laura,
Wong Brandon,
Dovichi Norman J.
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200209)23:18<3136::aid-elps3136>3.0.co;2-s
Subject(s) - chemistry , sodium dodecyl sulfate , chromatography , gel electrophoresis , laser induced fluorescence , fluorescence , fractionation , population , capillary electrophoresis , biochemistry , physics , demography , quantum mechanics , sociology
Capillary sodium dodecyl sulfate (SDS)‐DALT electrophoresis (SDS‐DALT‐CE) refers to CE separation of proteins based on their size; DALT is the abbreviation for Dalton, the unit used to describe molecular weight. In this work, seven proteins from 18 to 116 kDa were denatured by SDS, labeled by 3‐(2‐furoyl) quinoline‐2‐carboxaldehyde, separated by SDS‐DALT‐CE in polyethylene oxide sieving matrix, and detected by laser‐induced fluorescence (LIF) in a sheath flow cuvette. This method was combined with detergent differential fractionation, which is a protein fractionation method using a series of detergent‐containing buffers to sequentially extract protein fractions from cells, to analyze the proteins in HT29 human colon adenocarcinoma cells. In addition, on‐column labeling was demonstrated for protein analysis by SDS‐DALT‐CE with LIF, and applied to analysis of proteins in a single HT29 cancer cell. Most proteins had molecular masses from 10 to 120 kDa. Similar protein profiles were obtained for single cells and protein extract of a large cell population.