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Published on: September 3, 2014
Preparative Aspects of Immobilized pH Gradients
1Faculty of Pharmacy and Department of Biomedical Sciences and Technologies, University of Milano, Milano, Italy.
Abstract:
We will give here guidelines on the preparative aspects of immobilized pH gradients (IPGs), the latest and most powereful fractionation technique in electrophoresis. The load ability of IPG gels has been demonstrated to be at least 10 times higher than in conventional isoelectric focusing (IEF), thus approaching or even passing the load limit of isotachophoresis (1). The following aspects will be illustrated: (a) explorative runs aimed at defining the load capacity; (b) optimization of experimental parameters, such as ionic strength (I), pH gradient width, and gel thickness; (c) general considerations on the load ability and ionic strength of the milieu; (d) protein load as a function of %T in the matrix.
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