Isoelectric focusing by free solutioncapillary electrophoresis

Shiaw Min Chen, John E. Wiktorowicz

Research output: Contribution to journalArticle

79 Citations (Scopus)

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 - 1992
Externally publishedYes

Fingerprint

Ribonuclease T1
Capillary electrophoresis
Capillary Electrophoresis
Isoelectric Focusing
Electrophoresis
Proton-Motive Force
Isoelectric Point
Proteins
Experiments
Bacillus amyloliquefaciens ribonuclease

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Isoelectric focusing by free solutioncapillary electrophoresis. / Chen, Shiaw Min; Wiktorowicz, John E.

In: Analytical Biochemistry, Vol. 206, No. 1, 1992, p. 84-90.

Research output: Contribution to journalArticle

Chen, Shiaw Min ; Wiktorowicz, John E. / Isoelectric focusing by free solutioncapillary electrophoresis. In: Analytical Biochemistry. 1992 ; Vol. 206, No. 1. pp. 84-90.
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