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  Increased proteome coverage by combining PAGE and peptide isoelectric focusing: Comparative study of gel-based separation approaches.

Atanassov., I., & Urlaub, H. (2013). Increased proteome coverage by combining PAGE and peptide isoelectric focusing: Comparative study of gel-based separation approaches. Proteomics, 13(20), 2947-2955. doi:10.1002/pmic.201300035.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-7925-E Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-CB69-4
Genre: Journal Article

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Atanassov., I.1, Author              
Urlaub, H.1, Author              
Affiliations:
1Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              

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 Abstract: The in-depth analysis of complex proteome samples requires fractionation of the sample into subsamples prior to LC-MS/MS in shotgun proteomics experiments. We have established a 3D workflow for shotgun proteomics that relies on protein separation by 1D PAGE, gel fractionation, trypsin digestion, and peptide separation by in-gel IEF, prior to RP-HPLC-MS/MS. Our results show that applying peptide IEF can significantly increase the number of proteins identified from PAGE subfractionation. This method delivers deeper proteome coverage and provides a large degree of flexibility in experimentally approaching highly complex mixtures by still relying on protein separation according to molecular weight in the first dimension.

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Language(s): eng - English
 Dates: 2013-10-082013-10
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1002/pmic.201300035
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Title: Proteomics
Source Genre: Journal
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Pages: - Volume / Issue: 13 (20) Sequence Number: - Start / End Page: 2947 - 2955 Identifier: -