Picking Vanished Proteins from the Void: How to Collect and Ship/Share Extremely Dilute Proteins in a Reproducible and Highly Efficient Manner
Autor: | Andreas Otto, Michael Hecker, Knut Büttner, Dörte Becher, Jürgen Bartel, Florian Bonn |
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Rok vydání: | 2014 |
Předmět: |
Chromatography
Chemistry Solid Phase Extraction Proteins Reproducibility of Results Limiting Mass spectrometry Tandem mass spectrometry Mass spectrometric Analytical Chemistry Highly sensitive Tandem Mass Spectrometry Proteome Electrophoresis Polyacrylamide Gel Sample preparation Solid phase extraction Chromatography Liquid |
Zdroj: | Analytical Chemistry. 86:7421-7427 |
ISSN: | 1520-6882 0003-2700 |
Popis: | Successful proteome analyses of highly dilute samples are strongly dependent on optimized workflows considering especially sample preparation prior to highly sensitive mass spectrometric analysis. Various methods are available for enrichment of proteome samples, each characterized by specific advantages and disadvantages limiting their general application as a method of choice. Here we suggest an optimized universal protocol ensuring reproducibility and effective enrichment of dilute samples by commercial affinity beads. By comparably assessing the performance of the new protocol with selected standard enrichment techniques, we show the seamless application of the enrichment in common mass spectrometry based proteomic workflows. Further, novel applications are suggested including a facile storage and shipping of desiccated, trapped proteome samples at ambient temperatures and usage of the affinity beads for gel-free proteomic approaches. |
Databáze: | OpenAIRE |
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