Isoelectric Focusing

Maryvonne DR Brasher and Robin Thorpe, Division of Immunobiology, National Institute for Biological Standards and Control, Hertfordshire, UK

Isoelectric focusing (IEF) is an electrophoretic procedure capable of extremely high resolving power. During IEF, zwitterionic molecules arc separated according to their isoelectric point (pi; defined as the 'pH at which a zwitterion possesses no overall charge'), and the technique can therefore be used to analyze the composition of a mixture and/or to determine the precise pi of a component under investigation. Although usually applied to proteins or protein-containing extracts, the method can, at least theoretically, be used for analysis of any zwitterionic species. Methodology has been developed to allow IEF to be carried out on analytical and preparative scales.

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