Automated ultra-thin-layer SDS gel electrophoresis of proteins using noncovalent fluorescent labeling

Z. Csapó, Arpad Gerstner, M. Sasvári, A. Guttman

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Ultra-thin-layer SDS gel electrophoresis in conjunction with automated laser-induced fluorescence detection is a novel and powerful method for the analysis of fluorophore-labeled proteins. The technique described in this paper employs instant, noncovalent fluorophore labeling by the addition of a fluorescent staining dye to the sample proteins either during or immediately prior to the sample loading process. Thus, the method does not require time- consuming post- or pre-separation staining/labeling. By combining the multilane format of SDS polyacrylamide slab gel electrophoresis and the high separation efficiency of capillary SDS gel electrophoresis, Ultra-thin-layer SDS gel electrophoresis features rapid, high-throughput, and high-resolution analysis of proteins in the molecular mass range of 14-116 kDa. The good heat dissipation inherent to the ultrathin format enables the use of agarose and agarose-based composite separation matrixes, which can be easily replaced within the separation platform. Labeling efficiency as a function of the concentration of the staining dye, SDS, and proteins is thoroughly discussed. Detection sensitivity of the method was found to be at the low-femtomole level (1.25 ng/band), determined by analyzing a set of serial dilutions of standard proteins. Practical example of molecular mass determination and characterization of a complex protein mixture are also shown.

Original languageEnglish
Pages (from-to)2519-2525
Number of pages7
JournalAnalytical Chemistry
Volume72
Issue number11
DOIs
Publication statusPublished - Jun 1 2000

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Electrophoresis
Labeling
Gels
Proteins
Fluorophores
Molecular mass
Sepharose
Heat losses
Fluorescent Dyes
Dilution
Coloring Agents
Fluorescence
Throughput
Lasers
Composite materials

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Automated ultra-thin-layer SDS gel electrophoresis of proteins using noncovalent fluorescent labeling. / Csapó, Z.; Gerstner, Arpad; Sasvári, M.; Guttman, A.

In: Analytical Chemistry, Vol. 72, No. 11, 01.06.2000, p. 2519-2525.

Research output: Contribution to journalArticle

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