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The Alpher, Bethe and Gamow of IEF, the alpha‐Centaury of electrokinetic methodologies. Part II: Immobilized pH gradients
Author(s) -
Righetti Pier Giorgio
Publication year - 2007
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.200600256
Subject(s) - electrokinetic phenomena , chemistry , polyacrylamide , immobilized ph gradient , chromatography , electrophoresis , membrane , isoelectric focusing , combinatorial chemistry , computer science , analytical chemistry (journal) , biochemistry , polymer chemistry , enzyme
The present review (a follow up of Electrophoresis 2006, 27 , 923–938 on conventional IEF) highlights the developmental steps of the IPG technology, from a nebulous start limiting the technique to just 1 pH unit intervals up to the description of extended pH gradients, encompassing as much as 8.5 pH units. Although computer algorithms had been developed for optimizing recipes so as to obtain the most precise and most linear pH gradients, it was also realized that nonlinear pH intervals, covering the pH 3–10 range, would be extremely beneficial in 2‐D map analysis, since they would follow the p I distribution of proteins in living systems. The synthesis of a number of Immobiline chemicals (the acrylamido weak acids and bases meant to be incorporated into the nascent polyacrylamide chains) is also reported. The review ends with preparative aspects of IPGs, with the introduction of multicompartment electrolyzers with Immobiline membranes.