Sample preparation for mass spectrometry-based tissue (phospho)proteomics
Sander, S.; Bayramoglu, I.; Stumpe, M.; Restivo, G.; Levesque, M.; Dengjel, J.
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This protocol describes the workflow for the preparation of tissue samples for proteome and phosphoproteome analyses using mass spectrometry. The tissue samples are cryogenically pulverized and homogenized in a sucrose-based buffer to ensure proper tissue disruption. For depletion of lipid contaminants, proteins are purified using chloroform-methanol precipitation, followed by a resuspension in a urea-based buffer for enzymatic digestion. Peptides are desalted and enriched for phosphopeptides prior LC-MS/MS analysis. The workflow was developed for skin biopsies but is compatible with a broad range of tissue types. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=51 SRC="FIGDIR/small/732915v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@6a44aforg.highwire.dtl.DTLVardef@c34dc9org.highwire.dtl.DTLVardef@27d7ecorg.highwire.dtl.DTLVardef@1d1038e_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical abstractC_FLOATNO C_FIG
Matching journals
The top 3 journals account for 50% of the predicted probability mass.