Capillary sodium dodecyl sulfate-DALT electrophoresis with laser-induced fluorescence detection for size-based analysis of proteins in human colon cancer cells.
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.
Hu
Shen
S
Department of Chemistry, University of Washington, Seattle, WA 98195, USA.
Jiang
Jiang
J
Cook
Lillian M
LM
Richards
Dawn P
DP
Horlick
Laura
L
Wong
Brandon
B
Dovichi
Norman J
NJ
D000230:Adenocarcinoma / Q000737:chemistry; D003110:Colonic Neoplasms / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000706:statistics & numerical data; D005453:Fluorescence; D006801:Humans; D007834:Lasers; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D012967:Sodium Dodecyl Sulfate; D014407:Tumor Cells, Cultured
D009363:Neoplasm Proteins; D012967:Sodium Dodecyl Sulfate