Metal affinity-based purification of a red fluorescent protein

Y. Rahimi, S. Shrestha, S. K. Deo

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

A red fluorescent protein, DsRed, which emits fluorescence in the red region of the spectrum has become a popular alternative to green fluorescent protein as a label in biochemical and bioanalytical applications. In this study, we have developed a simple purification method for DsRed variants utilizing their inherent copper binding property. A purification procedure was developed and optimized using immobilized copper ions yielding a single strong band corresponding to purified DsRed proteins on the SDS-PAGE gel. A purification efficiency of higher than 95% was achieved. A spectral analysis and copper binding study was performed to verify activity of the purified proteins. The development of this method allows DsRed to play a dual role as a fluorescent reporter protein and as a purification affinity tag for a target protein. This simpler approach of purification should expand the utility of DsRed.

Original languageEnglish (US)
Pages (from-to)429-433
Number of pages5
JournalChromatographia
Volume65
Issue number7-8
DOIs
StatePublished - Apr 1 2007
Externally publishedYes

Keywords

  • Affinity chromatography
  • DsRed
  • Purification

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

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