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

Zheru Zhang, Sergey Krylov, Edgar Arriaga, Robert Polakowski, Norman J. Dovichi

Research output: Contribution to journalArticle

106 Scopus citations

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 ~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 106 cells.

Original languageEnglish (US)
Pages (from-to)318-322
Number of pages5
JournalAnalytical Chemistry
Volume72
Issue number2
DOIs
StatePublished - Jan 15 2000

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