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 language | English (US) |
---|---|
Pages (from-to) | 429-433 |
Number of pages | 5 |
Journal | Chromatographia |
Volume | 65 |
Issue number | 7-8 |
DOIs | |
State | Published - Apr 2007 |
Externally published | Yes |
Keywords
- Affinity chromatography
- DsRed
- Purification
ASJC Scopus subject areas
- Analytical Chemistry
- Clinical Biochemistry