One-dimensional protein analysis of an HT29 human colon adenocarcinoma cell.
A single HT29 human colon adenocarcinoma cell was introduced into a fused-silica capillary and lysed, and the protein content was fluorescently labeled with the fluorogenic reagent 3-(2-furoyl)quinoline-2-carboxaldehyde. The labeled proteins were separated by capillary electrophoresis in a submicellar buffer and detected by laser-induced fluorescence in a postcolumn sheath-flow cuvette. Several dozen components were resolved. A number of experiments were done to verify that these components were proteins. Most components of the single-cell electropherogram had the same mobility as components present in the 30-100 kDa fraction of a protein extract prepared from the cell culture. One component was identified as a approximately 100 kDa protein by co-injecting the sample with purified protein obtained from an SDS-PAGE gel. Protein expression varied significantly between cells, but the average expression was consistent with that observed from a protein extract prepared from 10(6) cells.
Zhang
Z
Z
Department of Chemistry, University of Alberta, Edmonton, Canada.
Krylov
S
S
Arriaga
E A
EA
Polakowski
R
R
Dovichi
N J
NJ
D019075:Electrophoresis, Capillary; D019073:HT29 Cells; D006801:Humans; D007834:Lasers; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000302:isolation & purification; D013050:Spectrometry, Fluorescence
D009363:Neoplasm Proteins