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The formation of stable pH gradients with weak monovalent buffers for isoelectric focusing in free solution
Author(s) -
Mosher Richard A.,
Thormann Wolfgang,
Graham Aly,
Bier Milan
Publication year - 1985
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.1150061106
Subject(s) - isoelectric focusing , electrolyte , isoelectric point , chemistry , diffusion , chromatography , range (aeronautics) , stability (learning theory) , analytical chemistry (journal) , biological system , materials science , thermodynamics , computer science , physics , electrode , organic chemistry , machine learning , biology , composite material , enzyme
Two methods which utilize simple buffers for the generation of stable pH gradients useful for preparative isoelectric focusing are compared and contrasted. The first employs preformed gradients comprised of two simple buffers in density‐stabilized free solution. The stability of this system is analyzed theoretically and by computer simulation. These precast gradients are limited to two buffering components, subject to diffusion, and restricted to the neutral pH region. An experimental application is presented. The second method utilizes neutral membranes to isolate electrolyte reservoirs of constant composition from the separation column. It is shown by computer simulation that steady state gradients can be formed at any pH range with any number of components in such a system.