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  Minimal, encapsulated proteomic-sample processing applied to copy-number estimation in eukaryotic cells

Kulak, N. A., Pichler, G., Paron, I., Nagaraj, N., & Mann, M. (2014). Minimal, encapsulated proteomic-sample processing applied to copy-number estimation in eukaryotic cells. NATURE METHODS, 11(3), 319-324. doi:10.1038/NMETH.2834.

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 Creators:
Kulak, Nils A.1, Author              
Pichler, Garwin1, Author              
Paron, Igor1, Author              
Nagaraj, Nagarjuna1, Author              
Mann, Matthias1, Author              
Affiliations:
1Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565159              

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Free keywords: MASS-SPECTROMETRY; GLOBAL ANALYSIS; DIGESTION; YEAST; QUANTIFICATION; EXPRESSION; PROTEINS; IDENTIFICATION; PURIFICATION; ENRICHMENT
 Abstract: Mass spectrometry (MSMS)-based proteomics typically employs multistep sample-preparation workflows that are subject to sample contamination and loss. We report an in-StageTip method for performing sample processing, from cell lysis through elution of purified peptides, in a single, enclosed volume. This robust and scalable method largely eliminates contamination or loss. Peptides can be eluted in several fractions or in one step for single-run proteome analysis. In one day, we obtained the largest proteome coverage to date for budding and fission yeast, and found that protein copy numbers in these cells were highly correlated (R-2 = 0.78). Applying the in-StageTip method to quadruplicate measurements of a human cell line, we obtained copy-number estimates for 9,667 human proteins and observed excellent quantitative reproducibility between replicates (R-2 = 0.97). The in-StageTip method is straightforward and generally applicable in biological or clinical applications.

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Language(s): eng - English
 Dates: 2014-03
 Publication Status: Published in print
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000332086100028
DOI: 10.1038/NMETH.2834
 Degree: -

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Title: NATURE METHODS
Source Genre: Journal
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Publ. Info: MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND : NATURE PUBLISHING GROUP
Pages: - Volume / Issue: 11 (3) Sequence Number: - Start / End Page: 319 - 324 Identifier: ISSN: 1548-7091