Universal and rapid method for purification of GFP-like proteins by the ethanol extraction

Protein Expr Purif. 2009 May;65(1):108-13. doi: 10.1016/j.pep.2008.11.008. Epub 2008 Dec 3.


GFP-like fluorescent proteins (FPs) are crucial in biological and biomedical studies. The majority of FP purification techniques either include multiple time-consuming chromatography steps with a low yield of the desired product or require prior protein modification (addition of special tags). In the present work, we propose an alternative ethanol extraction-based technique previously used for GFP purification and then modified for diverse FPs originated from different sources. The following recombinant FPs were expressed using Escherichia coli M15 (pREP4) strain as a host transformed with pQE30 plasmid bearing one of the target FP genes: TagCFP, TagGFP, TagYFP, TagRFP, TurboGFP, TurboRFP, Dendra2, TurboFP602 and KillerRed. Despite their diversity, all tested recombinant FPs were successfully purified and yielded a highly homogeneous product. The method is easily scalable for purification of any amount of protein and requires no expensive reagents and equipment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ethanol / chemistry*
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / isolation & purification*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification


  • Recombinant Proteins
  • Green Fluorescent Proteins
  • Ethanol