Prefractionation of protein samples prior to two‐dimensional electrophoresis
作者:
Garry L. Corthals,
Mark P. Molloy,
Ben R. Herbert,
Keith L. Williams,
Andrew A. Gooley,
期刊:
ELECTROPHORESIS
(WILEY Available online 1997)
卷期:
Volume 18,
issue 3‐4
页码: 317-323
ISSN:0173-0835
年代: 1997
DOI:10.1002/elps.1150180304
出版商: Wiley Subscription Services, Inc., A Wiley Company
关键词: Prefractionation;Preparative electrophoresis;Native electrophoresis;Two‐dimensional polyacrylamide gel electrophoresis;Proteome
数据来源: WILEY
摘要:
AbstractThousands 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 spec
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