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Prefractionation of protein samples prior to two-dimensional electrophoresis

✍ Scribed by Garry L. Corthals; Mark P. Molloy; Ben R. Herbert; Prof. Keith L. Williams; Andrew A. Gooley

Publisher: John Wiley and Sons
Year: 1997
Tongue: English
Weight: 724 KB
Volume: 18
Category: Article
ISSN: 0173-0835
DOI: 10.1002/elps.1150180304

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✦ Synopsis

Abstract

Thousands of proteins may be visualised on a two‐dimensional (2‐D) gel, but only hundreds are present at levels sufficient for chemical analysis. Therefore, prefractionation of protein samples prior to 2‐D polyacrylamide gel electrophoresis (PAGE) will be important for the investigation of proteins that are present at sub‐picogram levels in physiological samples. We describe an approach to prefractionate protein samples prior to 2‐D PAGE using the Gradiflow, which is a new (preparative) electrokinetic membrane apparatus designed to fractionate proteins in a number of different ways. We have fractionated human serum under nonreducing conditions using the ‘reflux’ mode, in which proteins are fractionated according to their relative mobility under controlled electrophoretic conditions, where the current is periodically reversed. We describe how fractionation occurs and present examples of enrichment of specific proteins.

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