One-dimensional protein analysis of an HT29 human colon adenocarcinoma cell.

Abstract

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.

• Authors: Z Zhang, S Krylov, E A Arriaga, R Polakowski, N J Dovichi
• Journal: Analytical chemistry (Anal Chem) Vol. 72 Issue 2 Pg. 318-22 (Jan 15 2000) ISSN: 0003-2700 [Print] United States
• PMID: 10658325 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Neoplasm Proteins

Topics

• Electrophoresis, Capillary
• HT29 Cells
• Humans
• Lasers
• Neoplasm Proteins (chemistry, isolation & purification)
• Spectrometry, Fluorescence