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Conductivity properties of carrier ampholyte pH gradients in isoelectric focusing
Author(s) -
Stoyanov Alexander V.,
Das Champak,
Fredrickson Carl K.,
Fan Z. Hugh
Publication year - 2005
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.200406170
Subject(s) - isoelectric focusing , conductivity , chemistry , analytical chemistry (journal) , electric field , electrical resistivity and conductivity , chromatography , resolution (logic) , voltage , physics , computer science , biochemistry , quantum mechanics , artificial intelligence , enzyme
The conductivity properties of natural pH gradient created by carrier ampholytes were studied during the process of isoelectric focusing (IEF). IEF was performed in capillaries (10–30 mm long) or in microchips with the same channel length. A 10–30× reduction of the conductivity of the separation medium was observed during the establishment of pH gradient. Results obtained using different IEF voltages indicate that there is a nonlinear relationship between the conductivity of an established pH gradient and the applied electric field. Our theoretical analysis using a simplified model generated values that reasonably agree with the experimental data. In addition, we found that above a certain electric field (∼300 V/cm), resolution does not increase with the applied voltage as predicated; we observed band‐broadening and gel breakdown. The approach presented in this work can be used for optimization of the IEF separation and judicious selection of IEF conditions.