Universal sample preparation method for proteome analysis

Nat Methods. 2009 May;6(5):359-62. doi: 10.1038/nmeth.1322. Epub 2009 Apr 19.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analytic Sample Preparation Methods / methods*
  • Animals
  • Brain Chemistry
  • Endopeptidases / chemistry
  • HeLa Cells
  • Humans
  • Iodoacetamide / chemistry
  • Liver / chemistry
  • Mass Spectrometry / methods*
  • Mice
  • Mitochondria / chemistry
  • Proteome / analysis*
  • Proteome / chemistry
  • Proteomics / methods*
  • Sodium Dodecyl Sulfate / chemistry
  • Tandem Mass Spectrometry / methods
  • Ultrafiltration / methods
  • Urea / chemistry

Substances

  • Proteome
  • Sodium Dodecyl Sulfate
  • Urea
  • Endopeptidases
  • Iodoacetamide