The next generation in ITRAQ Labelling: 8 Plex The advent of mass spectrometry based tagging methods, in particular iTRAQ® Reagents, have permitted relative expression measurements of large sets of proteins with a high degree of automation. The isobaric nature of the tags allows the protein samples to be pooled post labeling without increasing the complexity of the of the MS analysis. Identical peptides labeled with the different iTRAQ® reagents exhibit the same parent ion in MS. Upon MS/MS fragmention of the parent ion, unique signature ions are generated which distinguish the individual samples and hence the relative abundance among the samples can be determined As this iTRAQ® reagent technology has become established and gained acceptance in relative protein analysis, there is a requirement to expand the scope of this technique to enable a higher degree of multiplexing. A new set of reagents is now available , 8 Plex, that doubles the number of states that can be compared from 4 to 8 using the same robust NHS chemistry and easy to use protocols as the iTRAQ® Reagent - 4 plex. This provides greater flexibility in the design of experiments. In this presentation, multiplexing Proteomic strategies using 8 Plex iTRAQ® Reagent are described in a time progression study of biomarkers of Lung Fibrosis using a Mouse Model. The biological relevance of the changes found were confirmed using alternative techniques to confirm the power of this approach in pulling out biologically meaningful results in the analysis

Date :Wednesday 2nd April 2008

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