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Distorted zones in isoelectric‐focusing systems
Author(s) -
Jonsson Mats
Publication year - 1980
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.1150010304
Subject(s) - perpendicular , isoelectric focusing , isoelectric point , electrolyte , planar , convection , power (physics) , chemistry , quartz , analytical chemistry (journal) , electrode , mechanics , materials science , optics , chromatography , geometry , physics , computer science , thermodynamics , composite material , biochemistry , mathematics , computer graphics (images) , enzyme
A maximum utilization of the high resolving power of isoelectric focusing requires that the focused zones are both planar and perpendicular to the direction of focusing. This paper reports on extensive investigations carried out with a rectangular quartz column with provisions for direct photography of zonal shapes. The experiments have been performed in density gradients, but the conclusions may be generalized to other stabilizing media. It is demonstrated that distorted zones are formed if a second pH gradient, perpendicular to the direction of focusing, originates at one end of the column. The generation of such a pH gradient may have different causes: erroneous loading of the column, too high an initial power input, presence of salts of weak electrolytes in the sample, or unsuitable positioning of the electrodes. The effects of the second pH gradient on the shapes of the focused carrier‐ampholyte and protein zones are interpreted on the basis of the mechanism creating the natural axial pH gradient. The stability of the disturbed system is discussed in terms of the diffusional, electrical, and convective mass flows of its components.