Abstract
We describe a method, filter-aided sample preparation (FASP), which combines the advantages of in-gel and in-solution digestion for mass spectrometry-based proteomics. We completely solubilized the proteome in sodium dodecyl sulfate, which we then exchanged by urea on a standard filtration device. Peptides eluted after digestion on the filter were pure, allowing single-run analyses of organelles and an unprecedented depth of proteome coverage.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Analytic Sample Preparation Methods / methods*
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Animals
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Brain Chemistry
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Endopeptidases / chemistry
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HeLa Cells
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Humans
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Iodoacetamide / chemistry
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Liver / chemistry
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Mass Spectrometry / methods*
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Mice
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Mitochondria / chemistry
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Proteome / analysis*
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Proteome / chemistry
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Proteomics / methods*
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Sodium Dodecyl Sulfate / chemistry
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Tandem Mass Spectrometry / methods
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Ultrafiltration / methods
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Urea / chemistry
Substances
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Proteome
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Sodium Dodecyl Sulfate
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Urea
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Endopeptidases
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Iodoacetamide