Ultrathin-layer sodium dodecyl sulfate gel electrophoresis of proteins: Effects of gel composition and temperature on the separation of sodium dodecyl sulfate-protein complexes

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Abstract

This paper discusses the effects of gel composition and separation temperature on the migration properties of fluorescein-5-isothiocyanate- labeled protein molecular mass markers (ranging from 20 100 to 205 000 Da) in automated ultrathin-layer sodium dodecyl sulfate (SDS) gel electrophoresis. The separation mechanism with the agarose and composite agarose - linear polyacrylamide, agarose - hydroxyethyl cellulose, and agarose - polyethylene oxide matrices were all found to comply with the Ogston sieving model in the molecular mass range of the protein molecules investigated. Our temperature studies revealed that electrophoretic separation of SDS protein complexes is an activated process and, in pure agarose and in composite agarose hydroxyethyl cellulose and agarose - polyethylene oxide matrices that the separation requires increasing activation energy as a function of the molecular mass of the separated proteins. On the other hand, when linear polyacrylamide was used as composite additive, the activation energy demand of the separation decreased with increasing solute molecular mass. The sensitivity of the laser-induced fluorescent detection of the automated ultrathin-layer electrophoresis system was evaluated by injecting a series of dilutions of the markers and was found to be less than 2.5 ng/band for the fluorophore-labeled protein.

Original languageEnglish
Pages (from-to)834-840
Number of pages7
JournalELECTROPHORESIS
Volume21
Issue number5
DOIs
Publication statusPublished - Apr 19 2000

Keywords

  • Composite agarose gels
  • Fluorescent detection
  • Proteins
  • Sodium dodecyl sulfate gel electrophoresis
  • Ultrathin layer

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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