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

S. Di Palma, D.E. Stange, M.L. van de Wetering, H. Clevers, A. Heck, S. Mohammed

Research output: Contribution to journal/periodicalArticleScientificpeer-review

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Abstract

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.
Original languageEnglish
Pages (from-to)3814-3819
JournalJournal of Proteome Research
Volume10
Issue number8
DOIs
Publication statusPublished - 2011

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