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Preparative isoelectric focusing in a cellulose‐based separation medium
Author(s) -
Šalplachta Jiří,
Horká Marie,
Šlais Karel
Publication year - 2017
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201700036
Subject(s) - chromatography , isoelectric focusing , cellulose , chemistry , analyte , ethylene glycol , electrophoresis , aqueous solution , capillary electrophoresis , pulmonary surfactant , isoelectric point , glycerol , organic chemistry , biochemistry , enzyme
An improved preparative method based on isoelectric focusing of analytes in a cellulose‐based separation medium is described in this study. Cellulose is suspended in an aqueous solution of simple buffers, ethylene glycol, glycerol, nonionic surfactant, and colored p I markers. Water partially evaporates during focusing run and the separation takes place in an in situ generated layer of cellulose, which has a gel‐like appearance at the end of analysis. Final positions of analytes are indicated by the positions of zones of focused p I markers. Fractions, segments of the separation medium with analytes, can be simply collected by spatula and analyzed by downstream analytical methods. Good focusing ability of the new method and almost quantitative recovery of model proteins, cytochrome c and bovine serum albumin, was verified by gel electrophoresis and capillary isoelectric focusing of the collected fractions.