Green fluorescent protein purification by organic extraction

Protein Expr Purif. 1998 Dec;14(3):382-6. doi: 10.1006/prep.1998.0981.

Abstract

Green fluorescent protein (GFP) is widely used as an excellent reporter molecule in biochemistry and cell biology. Some biochemical and immunological assays require high-purity GFP. However, the majority of current procedures for GFP purification include multiple time-consuming chromatography steps with a low yield of the desired product or require tag-containing proteins. An alternative method is described for the GFP purification without affinity extensions using organic extraction yielding a highly homogeneous protein indistinguishable in spectroscopic properties from that purified by previous methods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Butanol
  • Ammonium Sulfate
  • Chemical Fractionation
  • Chemical Precipitation
  • Chloroform
  • Chromatography, Liquid
  • Circular Dichroism
  • Escherichia coli / genetics
  • Ethanol
  • Green Fluorescent Proteins
  • Luminescent Proteins / genetics
  • Luminescent Proteins / isolation & purification*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification*
  • Sepharose / analogs & derivatives
  • Solvents
  • Spectrometry, Fluorescence
  • Water

Substances

  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Solvents
  • Water
  • Green Fluorescent Proteins
  • Ethanol
  • phenyl-sepharose
  • Chloroform
  • 1-Butanol
  • Sepharose
  • Ammonium Sulfate