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Isotachophoretic focusing on thin gel slab: A new and powerful electrophoretic method of protein analysis
Author(s) -
Charlionet Roland,
Bringard Aline,
Davrinche Christian,
Fontaine Marc
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.1150071205
Subject(s) - isotachophoresis , electrophoresis , chromatography , chemistry , stack (abstract data type) , electric field , computer science , electrolyte , physics , programming language , electrode , quantum mechanics
Abstract The theoretical principle and experimental verification of a new separation method for proteins in thin gel slabs called “isotachophoretic focusing” are presented. Isotachophoretic focusing is basically isotachophoresis with two new features: (i) By using adequate spacers a continuous mobility spectrum is generated. Under isotachophoresis conditions, these spacers give rise to a linear electric field gradient in which proteins can be separated in form of focusing bands. The synthesis and isolation of suitable spacers for isotachophoretic focusing are described. (ii) The stack is immobilized or, at least, its migration is retarded by exploiting the counterflow of electroendosmosis resulting from the action of the electric field on charged groups linked to the gel. Immobilization of the stack allows its formation to equilibrium and the protein separation process to continue as long as necessary. Details are given concerning the experimental set‐up and factors involved in the optimization of the technique. The separations obtained by isotachophoretic focusing are compared with those obtained by classical electrophoresis.