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Affinity chromatography and capillary electrophoresis for analysis of the yeast ribosomal proteins

Author(s): Miriam S. Goyder | Keith R. Willison | David R. Klug | Andrew J. deMello | Oscar Ces

Journal: BMB Reports
ISSN 1976-6696

Volume: 45;
Issue: 4;
Start page: 233;
Date: 2012;
Original page

Keywords: Affinity chromatography | Capillary electrophoresis | EVV 2DIR | Proteomics | Ribosomal proteins

We present a top down separation platform for yeast ribosomalproteins using affinity chromatography and capillary electrophoresiswhich is designed to allow deposition of proteins ontoa substrate. FLAG tagged ribosomes were affinity purified, andrRNA acid precipitation was performed on the ribosomes followedby capillary electrophoresis to separate the ribosomalproteins. Over 26 peaks were detected with excellent reproducibility(<0.5% RSD migration time). This is the first reportedseparation of eukaryotic ribosomal proteins using capillaryelectrophoresis. The two stages in this workflow, affinity chromatographyand capillary electrophoresis, share the advantagesthat they are fast, flexible and have small sample requirementsin comparison to more commonly used techniques. This methodis a remarkably quick route from cell to separation that hasthe potential to be coupled to high throughput readout platformsfor studies of the ribosomal proteome. [BMB reports2012; 45(4): 233-238]

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