Universal sample preparation method for proteome analysis.
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.
Wiśniewski
Jacek R
JR
Department of Proteomics and Signal Transduction, Max Planck Institute for Biochemistry, Martinsried, Germany. jwisniew@biochem.mpg.de
Zougman
Alexandre
A
Nagaraj
Nagarjuna
N
Mann
Matthias
M
D053000:Analytic Sample Preparation Methods / Q000379:methods*; D000818:Animals; D001923:Brain Chemistry; D010450:Endopeptidases / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D007460:Iodoacetamide / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008928:Mitochondria / Q000737:chemistry; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods; D014462:Ultrafiltration / Q000379:methods; D014508:Urea / Q000737:chemistry
D020543:Proteome; D012967:Sodium Dodecyl Sulfate; D014508:Urea; D010450:Endopeptidases; D007460:Iodoacetamide