Size-Dependent Separation of Proteins Denatured in SDS by Capillary Electrophoresis Using a Replaceable Sieving Matrix

W. E. Werner, D. M. Demorest, J. Stevens, J. E. Wiktorowicz

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

The determination of molecular weights by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is one of the most powerful electrophoretic techniques for protein characterization. A separation media has been developed which allows this type of analysis to be performed in the capillary format. A replaceable polymeric solution rather than a polymerized gel was used as the sieving matrix. This solution allowed for the separation of proteins denatured in SDS in a size-dependent manner as demonstrated by the linear correlation between the proteins′ relative migration and the concentration of the sieving matrix (Ferguson plot). The logarithm of the molecular weight of protein standards correlated linearly with the relative mobility of the denatured proteins over the molecular weight range of 14,000 to 205,000. The calculated resolution at half-peak height was such that proteins that differed by as little as 4% in molecular weight would be resolved. Finally, the integrated peak areas at 215 nm were linearly proportional to the mass of the protein injected.

Original languageEnglish (US)
Pages (from-to)253-258
Number of pages6
JournalAnalytical Biochemistry
Volume212
Issue number1
DOIs
StatePublished - Jul 1993
Externally publishedYes

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Capillary electrophoresis
Capillary Electrophoresis
Molecular Weight
Molecular weight
Proteins
Electrophoresis
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Gels

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

Size-Dependent Separation of Proteins Denatured in SDS by Capillary Electrophoresis Using a Replaceable Sieving Matrix. / Werner, W. E.; Demorest, D. M.; Stevens, J.; Wiktorowicz, J. E.

In: Analytical Biochemistry, Vol. 212, No. 1, 07.1993, p. 253-258.

Research output: Contribution to journalArticle

Werner, W. E. ; Demorest, D. M. ; Stevens, J. ; Wiktorowicz, J. E. / Size-Dependent Separation of Proteins Denatured in SDS by Capillary Electrophoresis Using a Replaceable Sieving Matrix. In: Analytical Biochemistry. 1993 ; Vol. 212, No. 1. pp. 253-258.
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