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Diagonal isoelectric focusing on immobilized pH gradients: Concept and preliminary performance tests
Author(s) -
Fawcett John S.,
Chrambach Andreas
Publication year - 1986
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.1150070402
Subject(s) - isoelectric focusing , immobilized ph gradient , isoelectric point , diagonal , range (aeronautics) , analytical chemistry (journal) , field strength , chemistry , conductance , chromatography , electric field , electric field gradient , field (mathematics) , resolution (logic) , materials science , physics , mathematics , magnetic field , computer science , composite material , geometry , condensed matter physics , biochemistry , quantum mechanics , artificial intelligence , pure mathematics , enzyme
Abstract Electrofocusing on diagonal immobilized pH gradients was designed to combine high field strength with a flat pH gradient over a wide pH range, circumventing the need for extremely high voltage in long, conventionally oriented focusing gels. The technique was found to be applicable at 500 V cm −1 to gels of 35 cm length of a flat pH gradient with a predicted pH range of 3.8 to 9.8, allowing for a focusing time of 2 h. Note that the equivalent flat pH gradient gel of 25 cm length at the same field strength would require application of 12 500 V. However, preliminary results indicate that the theoretically expected improvement in resolution due to high field strength does not materialize. This may be due to a distorted electric field caused by non‐uniform conductance features inherent in the method and by the passage of the current through the isoelectric protein zones.