Green fluorescent protein purification by organic extraction.
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.
Yakhnin
A V
AV
Branch of the Institute of Bioorganic Chemistry RAS, Pushchino, Moscow Region, 142292, Russia. yak@fibkh.serpukhov.su
Vinokurov
L M
LM
Surin
A K
AK
Alakhov
Y B
YB
D020001:1-Butanol; D000645:Ammonium Sulfate; D005591:Chemical Fractionation; D011232:Chemical Precipitation; D002725:Chloroform; D002853:Chromatography, Liquid; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics; D000431:Ethanol; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012685:Sepharose / Q000031:analogs & derivatives; D012997:Solvents; D013050:Spectrometry, Fluorescence; D014867:Water
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D012997:Solvents; D014867:Water; D049452:Green Fluorescent Proteins; D000431:Ethanol; C020927:phenyl-sepharose; D002725:Chloroform; D020001:1-Butanol; D012685:Sepharose; D000645:Ammonium Sulfate