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Highly sensitive proteome analysis of FACS-sorted adult colon stem cells

Palma Di, S. and Stange, D.E. and Wetering van de, M.L. and Clevers, H. and Heck, A. and Mohammed, S. (2011) Highly sensitive proteome analysis of FACS-sorted adult colon stem cells. Journal of Proteome Research, 10, 3814-9. ISSN 1535-3893.

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In proteomics, multidimensional liquid chromatography combined with mass spectrometry has become a standard technique to reduce sample complexity and tackle the vast dynamic range. Such fractionation is necessary to obtain a comprehensive analysis of biological samples such as tissues and cell lines. However, extensive fractionation comes at the expense of sample losses, hampering the analysis of limited material. We previously described a highly sensitive multidimensional chromatographic strategy based on a combination of hydrophilic interaction liquid chromatography and reversed phase chromatography, which allows proteomic analysis with minimal sample losses. Here we apply this strategy to the analysis of a limited number of FACS-sorted colon stem cells extracted from mouse intestine, obtaining a proteome coverage comparable to current methods that generally require 100-fold more starting material. We propose that this alternative multidimensional chromatographic technology will find ample application such as in the analysis of distinct cellular populations obtained by laser microdissection.

Item Type:Article
Institutes:Hubrecht Instituut
ID Code:12647
Deposited On:19 Sep 2012 12:46
Last Modified:14 Oct 2012 19:05

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