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.
Cite this paper
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