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Solution phase isoelectric fractionation in the multi-compartment electrolyser: a divide and conquer strategy for the analysis of complex proteomes.
Authors:Gary B Smejkal  Alexander Lazarev
Institution:Proteome Systems, 14 Gill Street, Woburn, MA 01801, USA. smejkal@proteomesystems.com
Abstract:Sample complexity frequently interferes with the analysis of low-abundance proteins by two-dimensional gel electrophoresis (2DGE). Ideally, high abundance proteins should be removed, allowing low-abundance proteins to be applied at much higher concentrations than is possible with the unfractionated sample. One approach is to partition the sample in a manner that segregates the bulk of extraneous proteins from the protein(s) of interest. Solution phase isoelectric focusing in the multi-compartment electrolyser generates fractions of discrete isoelectric point (pI) intervals allowing isolated narrow segments of a proteome to be analysed individually by 2DGE. It is particularly useful for the isolation of low-abundance proteins of extremely basic or acidic pI.
Keywords:immobilised pH gradient  multi-compartment electrolyser  proteomics  solution phase isoelectric focusing  two-dimensional gel electrophoresis
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