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Mass spectrometric analysis of the electroeluates of fluorescent proteins after preparative electrophoresis in the automated HPGE‐1000 apparatus
Author(s) -
Yarmola Elena,
Chrambach Andreas,
Nguyen Viet Quoc,
Yergey Alfred L.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990301)20:3<445::aid-elps445>3.0.co;2-j
Subject(s) - electroelution , chromatography , fluorescence , mass spectrometry , chemistry , electrophoresis , green fluorescent protein , gel electrophoresis , analytical chemistry (journal) , polyacrylamide gel electrophoresis , biochemistry , physics , quantum mechanics , gene , enzyme
Bands of green fluorescent protein (GFP) and R‐phycoerythrin (PHYCO) in gel electrophoresis on the automated apparatus for gel electrophoresis with periodic fluorescence scanning (HPGE), the HPGE‐1000 apparatus, were retrieved from the gel by electroelution. While PHYCO was recovered in a single volume of electroeluate buffer after the predicted migration time, GFP fluorescence was lost under the same conditions and could only be recovered using multiple changes of electroeluate buffer. The multiple volumes of buffer necessitated pooling, concentration, and storage, conditions under which a minor GFP component, GFP‐II, formed artifactually. PHYCO after electroelution also exhibits a minor component present in the original preparation. The electroeluate of GFP, transferred into a mass spectrometer after pooling, concentration and storage, is indistinguishable in mass from the original preparation.