Micellar electrokinetic chromatography with diode laser-induced fluorescence detection as a tool for investigating the fluorescence labeling of proteins

Peng Jing, Takashi Kaneta, Totaro Imasaka

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

We describe the use of micellar electrokinetic chromatography with diode laser-induced fluorometry (MEKC-DLIF) as a tool to characterize a labeling reaction between sulfoindocyanine succinimidyl ester (Cy5), a cyanine fluorescent dye, and a model protein, bovine serum albumin (BSA). To decrease the influence of imprecise injection, methylene blue was added as an internal standard. Labeled BSA was completely separated from the unconjugated Cy5, and methylene blue under optimized conditions. A kinetic study of the reaction was performed by changing some parameters, such as reaction buffer pH, reaction buffer concentration and the initial concentration of BSA. A comparison between the current method and traditional methods was made.

Original languageEnglish
Pages (from-to)550-555
Number of pages6
JournalElectrophoresis
Volume23
Issue number4
DOIs
Publication statusPublished - Mar 27 2002
Externally publishedYes

Keywords

  • Bioconjugate chemistry
  • Diode laser
  • Dye characterization
  • Micellar electrokinetic chromatography
  • Protein

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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