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TMT (Tandem Mass Tags) and iTRAQ (Isobaric Tag for Relative Absolute Quantitation) are peptide-isotope-labeling techniques developed by AB Sciex and Thermo, respectively. These techniques can use up to 16 stable isotope tags (TMT) for different samples.
The iTRAQ/TMT reagent is composed of three components: the reporter group, the balance group, and the reactive group. The reactive groups form 2-16 isotope labels with equal masses. The iTRAQ/TMT reagent labels enzymatic peptides and can react with primary amine groups at the N-terminus and side chains of amino acids. In the primary spectra, the same peptide from different samples is labeled with the same mass-to-charge. In the secondary spectra, the reporter group produces the different intensity of reports ions with different isotope tags. The quantitative information of the same peptide between different samples can be obtained according to the abundance of the reporter ions, and then the quantitative information of the protein is obtained by software processing. The basic principle and technical flow are illustrated in the figure below:
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