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Native and sodium dodecyl sulfate‐capillary gel electrophoresis of proteins on a single microchip
Author(s) -
Tsai ShuoWen,
Loughran Michael,
Suzuki Hiroaki,
Karube Isao
Publication year - 2004
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/elps.200305725
Subject(s) - capillary electrophoresis , electrophoresis , sodium dodecyl sulfate , chromatography , chemistry , stacking , analytical chemistry (journal) , gel electrophoresis , gel electrophoresis of nucleic acids , electrode , free flow electrophoresis , cathode , capillary action , anode , gel electrophoresis of proteins , materials science , polyacrylamide gel electrophoresis , biochemistry , composite material , organic chemistry , enzyme
Simultaneous electrophoresis of both native and Sodium dodecyl sulfate (SDS) proteins was observed on a single microchip within 20 min. The capillary array prevented lateral diffusion of SDS components and avoided cross contamination of native protein samples. The planar sputtered electrode format provided a more uniform distribution of separation voltage into each of the 36 parallel microchannel capillaries than platinum wire electrodes commonly used in conventional electrophoresis. The customized geometry of the stacking capillary machined into the cover plate of the microchip facilitated reproducible sample injection without the requirement for stacking gel. Polyimide served as a mask and facilitated insulation of the anode and cathode to prevent electrode lift off and deterioration during continuous electrophoresis, even at a constant current of 8 mA. Improved protein separation was observed during capillary electrophoresis at lower currents. Ferguson plot analysis confirmed the electrophoretic mobility of native globular proteins in accordance with their charge and size. Corresponding Ferguson plot analysis of SDS‐associated proteins on the same chip confirmed separation of marker proteins according to their molecular weight.