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Parallel post-source decay for increasing protein identification confidence levels from 2-D gels
Department of Protein Technology, Lund University, Lund, Sweden.
Department of Biological Chemistry, University of Padova, Padova, Italy.
Waters Corporation Facilities, Manchester, UK.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm , Ludwig Institute for Cancer Research.
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2008 (English)In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 8, no 9, 1771-1779 p.Article in journal (Refereed) Published
Abstract [en]

Peptide mass fingerprinting (PMF) has over the years become one of the most commonly used tools for high-throughput analysis and identification of proteins. This method is applicable when relatively simple samples have to be analysed and it is commonly used for analysing proteins previously separated by 2-DE. The most common type of instrument used for this approach is the MALDI-TOF that has proved to be particularly suitable for the PMF analysis because of its characteristics of speed, robustness, sensitivity and automation. We have used a MALDI-TOF equipped with a novel parallel PSD capability (MALDI micro MX), to perform the analysis of two sets of different biological samples isolated by 2-DE. By using a method that integrates the data obtained by PMF analysis with the PSD data obtained in the same experiment, we show that the new multiplexed PSD solution increases the protein identification rate compared to the normal PMF approach. We also investigated the use of a charge-directed fragmentation modification reagent to improve the identification rate and confidence levels.

Place, publisher, year, edition, pages
2008. Vol. 8, no 9, 1771-1779 p.
Keyword [en]
Confidence level, Database searching, Peptide mass fingerprinting, Post-source decay
National Category
Medical and Health Sciences
URN: urn:nbn:se:uu:diva-17585DOI: 10.1002/pmic.200700894ISI: 000255720600007PubMedID: 18442167OAI: oai:DiVA.org:uu-17585DiVA: diva2:45356
Available from: 2008-07-10 Created: 2008-07-10 Last updated: 2011-03-18Bibliographically approved

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Hellman, Ulf
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