Validation of an Easy Acetonitrile Fractionation for the Simplification of Protein Samples Prior to Proteomics Analyses

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DOI: 10.4236/oalib.1101928    869 Downloads   1,505 Views  

ABSTRACT

In most proteomics analyses and in particular for the “off-gel” approaches, based essentially on chromatography, the complexity of the proteome should be reduced; otherwise identifications can be hindered, especially if the mass spectrometry analysis is not conducted using state-of-the-art instrumentation. Even if the method used is a bottom-up proteomics, it appears mandatory to pre-fractionate the proteins in order to reduce the complexity. We report here the development and validation of a pre-fractionation based on the differential solubilisation of proteins using increasing concentrations of acetonitrile (ACN). This “ACN fractionation” was applied to the study of the Triton X-100 soluble sub-proteome of brain capillary endothelial cells (BCEC) with re-induced blood-brain barrier (BBB) functions.

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Karamanos, Y. , Deracinois, B. , Pottiez, G. and Duban-Deweer, S. (2015) Validation of an Easy Acetonitrile Fractionation for the Simplification of Protein Samples Prior to Proteomics Analyses. Open Access Library Journal, 2, 1-6. doi: 10.4236/oalib.1101928.

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