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Topics {✒️}

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Questions {❓}

• An insight into iTRAQ: where do we stand now?
• An insight into iTRAQ: where do we stand now?

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         headline:An insight into iTRAQ: where do we stand now?
         description:The iTRAQ (isobaric tags for relative and absolute quantification) technique is widely employed in proteomic workflows requiring relative quantification. Here, we review the iTRAQ literature; in particular, we focus on iTRAQ usage in relation to other commonly used quantitative techniques e.g. stable isotope labelling in culture (SILAC), label-free methods and selected reaction monitoring (SRM). As a result, we identify several issues arising with respect to iTRAQ. Perhaps frustratingly, iTRAQ’s attractiveness has been undermined by a number of technical and analytical limitations: it may not be truly quantitative, as the changes in abundance reported will generally be underestimated. We discuss weaknesses and strengths of iTRAQ as a methodology for relative quantification in the light of this and other technical issues. We focus on technical developments targeted at iTRAQ accuracy and precision, use of 4-plex over 8-plex reagents and application of iTRAQ to post-translational modification (PTM) workflows. We also discuss iTRAQ in relation to label-free approaches, to which iTRAQ is losing ground.
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            MS/MS instrumentation
            Isotopic impurities
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            Modelling
            PTMs
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      headline:An insight into iTRAQ: where do we stand now?
      description:The iTRAQ (isobaric tags for relative and absolute quantification) technique is widely employed in proteomic workflows requiring relative quantification. Here, we review the iTRAQ literature; in particular, we focus on iTRAQ usage in relation to other commonly used quantitative techniques e.g. stable isotope labelling in culture (SILAC), label-free methods and selected reaction monitoring (SRM). As a result, we identify several issues arising with respect to iTRAQ. Perhaps frustratingly, iTRAQ’s attractiveness has been undermined by a number of technical and analytical limitations: it may not be truly quantitative, as the changes in abundance reported will generally be underestimated. We discuss weaknesses and strengths of iTRAQ as a methodology for relative quantification in the light of this and other technical issues. We focus on technical developments targeted at iTRAQ accuracy and precision, use of 4-plex over 8-plex reagents and application of iTRAQ to post-translational modification (PTM) workflows. We also discuss iTRAQ in relation to label-free approaches, to which iTRAQ is losing ground.
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         iTRAQ
         Ratio underestimation
         Relative quantification
         MS/MS instrumentation
         Isotopic impurities
         Statistical validation
         Modelling
         PTMs
         Label-free
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         Characterization and Evaluation of Materials
         Food Science
         Monitoring/Environmental Analysis
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      name:Biological and Environmental Systems Group, The ChELSI Institute, Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK
      name:Biological and Environmental Systems Group, The ChELSI Institute, Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK
      name:Biological and Environmental Systems Group, The ChELSI Institute, Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK
      name:Biological and Environmental Systems Group, The ChELSI Institute, Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK
      name:Biological and Environmental Systems Group, The ChELSI Institute, Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK
      name:The ChELSI Institute, Department of Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK
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