Virtual 2-D Gel Electrophoresis - American Chemical Society

Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109-0606, and ... proteins; virtual 2-D gels found 250 proteins rang...
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Anal. Chem. 2001, 73, 4063-4070

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Virtual 2-D Gel Electrophoresis: Visualization and Analysis of the E. coli Proteome by Mass Spectrometry Rachel R. Ogorzalek Loo,*,†,‡ James D. Cavalcoli,§ Ruth A. VanBogelen,§ Charles Mitchell,†,| Joseph A. Loo,§ Brian Moldover,§,⊥ and Philip C. Andrews*,†

Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109-0606, and Pfizer Global Research and Development, 2800 Plymouth Road, Ann Arbor, Michigan 48105

Mass spectrometric surface analysis of isoelectric focusing gels provides an ultrasensitive approach to proteome analysis. This “virtual 2-D gel” approach, in which mass spectrometry is substituted for the size-based separation of SDS-PAGE, provides advantages in mass resolution and accuracy over classical 2-D gels and can be readily automated. Protein identities can be postulated from molecular mass ((0.1-0.2% for proteins of 2 to