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Comparison of one- and two-dimensional liquid chromatography approaches in the label-free quantitative analysis of methylocella silvestris
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Patel, Nisha A., Crombie, Andrew, Slade, Susan E., Thalassinos, Konstantinos, Hughes, Chris, Connolly, Joanne B., Langridge, James, Murrell, J. C. (J. Colin) and Scrivens, James H. (2012) Comparison of one- and two-dimensional liquid chromatography approaches in the label-free quantitative analysis of methylocella silvestris. Journal of Proteome Research, Vol.11 (No.9). pp. 4755-4763. doi:10.1021/pr300253s ISSN 1535-3893.
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Official URL: http://dx.doi.org/10.1021/pr300253s
Abstract
The proteome of the bacterium Methylocella silvestris has been characterized using reversed phase ultra high pressure liquid chromatography (UPLC) and two-dimensional reversed phase (high pH)–reversed phase (low pH) UPLC prior to mass spectrometric analysis. Variations in protein expression levels were identified with the aid of label-free quantification in a study of soluble protein extracts from the organism grown using methane, succinate, or propane as a substrate. The number of first dimensional fractionation steps has been varied for 2D analyses, and the impact on data throughput and quality has been demonstrated. Comparisons have been made regarding required experimental considerations including total loading of biological samples required, instrument time, and resulting data file sizes. The data obtained have been evaluated with respect to number of protein identifications, confidence of assignments, sequence coverage, relative levels of proteins, and dynamic range. Good qualitative and quantitativ...
| Item Type: | Journal Article | ||||
|---|---|---|---|---|---|
| Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
| Journal or Publication Title: | Journal of Proteome Research | ||||
| Publisher: | American Chemical Society | ||||
| ISSN: | 1535-3893 | ||||
| Official Date: | 2012 | ||||
| Dates: |
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| Volume: | Vol.11 | ||||
| Number: | No.9 | ||||
| Page Range: | pp. 4755-4763 | ||||
| DOI: | 10.1021/pr300253s | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published | ||||
| Access rights to Published version: | Restricted or Subscription Access |
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