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JXB Advance Access originally published online on March 30, 2006
Journal of Experimental Botany 2006 57(7):1501-1508; doi:10.1093/jxb/erj168
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

A perspective on the use of iTRAQTM reagent technology for protein complex and profiling studies

Lynn R Zieske*

Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404, USA

*E-mail: zieskelr{at}appliedbiosystems.com

Proteomic research includes the characterization of protein mixtures in order to understand complex biological systems and determine relationships between proteins, their function, and protein–protein interactions. Often the goal of such research is to monitor changes of proteins in perturbed systems, a type of study referred to as differential expression analysis. To perform these studies requires the ability to execute some type of differential comparison of a given protein state in reference to some type of a control. The iTRAQTM reagents are a set of isobaric reagents which are amine specific and allow for the identification and quantitation of up to four different samples simultaneously. The amine specificity of these reagents makes most peptides in a sample amenable to this labeling strategy with no loss of information from samples involving post-translational modifications, such as the scrutiny of signal transduction pathways that often involve phosphorylation phenomena. In addition, the multiplexing capacity of these reagents allows for information replication within certain LC-MS/MS experimental regimes, providing additional statistical validation within any given experiment. The results presented herein demonstrate a few examples of the wide variety of quantitative information that can be realized when undertaking such experimental approaches. These include temporal analysis of drug-induced-protein expression, discovery and elucidation of disease markers, and protein–protein interactions in multi-protein complexes.

Key words: Differential expression, iTRAQ reagents, protein profiling, stable isotope labelling


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