iTRAQ reagent-based quantitative proteomic analysis on a linear ion trap mass spectrometer.

iTRAQ reagent-based quantitative proteomic analysis on a linear ion trap mass spectrometer. Research Abstract Details 

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iTRAQ reagent-based quantitative proteomic analysis on a linear ion trap mass spectrometer. Abstract Text:

Timothy J Griffin,Hongwei Xie,Sricharan Bandhakavi,Jonathan Popko,Archana Mohan,John V Carlis,LeeAnn Higgins,Timothy J Griffin,Hongwei Xie,Sricharan Bandhakavi,Jonathan Popko,Archana Mohan,John V Carlis,LeeAnn Higgins,

For proteomic analysis using tandem mass spectrometry, linear ion trap instruments provide unsurpassed sensitivity but unreliably detect low mass peptide fragments, precluding their use with iTRAQ reagent-labeled samples. Although the popular LTQ linear ion trap supports analyzing iTRAQ reagent-labeled peptides via pulsed Q dissociation, PQD, its effectiveness remains questionable. Using a standard mixture, we found careful tuning of relative collision energy necessary for fragmenting iTRAQ reagent-labeled peptides, and increasing microscan acquisition and repeat count improves quantification but identifies somewhat fewer peptides. We developed software to calculate abundance ratios via summing reporter ion intensities across spectra matching to each protein, thereby providing maximized accuracy. Testing found that results closely corresponded between analysis using optimized LTQ-PQD settings plus our software and using a Qstar instrument. Thus, we demonstrate the effectiveness of LTQ-PQD analyzing iTRAQ reagent-labeled peptides, and provide guidelines for successful quantitative proteomic studies.

iTRAQ reagent-based quantitative proteomic analysis on a linear ion trap mass spectrometer. Publishing Authors By Initials

TJ Griffin,H Xie,S Bandhakavi,J Popko,A Mohan,JV Carlis,L Higgins,TJ Griffin,H Xie,S Bandhakavi,J Popko,A Mohan,JV Carlis,L Higgins,

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iTRAQ reagent-based quantitative proteomic analysis on a linear ion trap mass spectrometer. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of proteome research

VOLUME: 6

Page Numbers: 4200-9

Journal Abbreviation: J. Proteome Res.

ISSN: 1535-3893

DAY: 29

MONTH: 09

YEAR: 2007

iTRAQ reagent-based quantitative proteomic analysis on a linear ion trap mass spectrometer. Information

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LANGUAGE: eng

NlmUniqueID: 101128775

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Grant and Affiliation Information for iTRAQ reagent-based quantitative proteomic analysis on a linear ion trap mass spectrometer.

AFFILIATION: Department of Biochemistry, Molecular Biology, and Biophysics, Center for Mass Spectrometry and Proteomics, University of Minnesota, Minneapolis, Minnesota 55455, USA. tgriffin@umn.edu

Country: United States

AGENCY: United States NIDDK

GRANT: DK073731

ACRONYM: DK

MEDLINETA: J Proteome Res

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