Isoelectric focusing by free solutioncapillary electrophoresis

Shiaw Min Chen, John E. Wiktorowicz

    Research output: Contribution to journalArticle

    80 Scopus citations

    Abstract

    A reproducible, quantitative isoelectric focusing method using capillary electrophoresis that exhibits high resolution and linearity over a wide pH gradient was developed. RNase T1 and RNase ba are two proteins that have isoelectric points (pI's) at the two extremes of a pH 3-10 gradient. Site-directed mutants of the former were separated from the wild-type form and pI's determined in the same experiment. The pI's of RNase T1 wild-type, its three mutants, and RNase ba were determined for the first time as 2.9, 3.1, 3.1, 3.3, and 9.0, respectively. The paper describes the protocol for isoelectric focusing by capillary electrophoresis, as well as presenting data describing the linearity, resolution, limits of mass loading, and reproducibility of the method.

    Original languageEnglish (US)
    Pages (from-to)84-90
    Number of pages7
    JournalAnalytical Biochemistry
    Volume206
    Issue number1
    DOIs
    StatePublished - Oct 1992

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    ASJC Scopus subject areas

    • Biophysics
    • Biochemistry
    • Molecular Biology
    • Cell Biology

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