Why can't TMT and iTRAQ be used simultaneously?
Both iTRAQ (Isobaric Tag for Relative Absolute Quantitation) and TMT (Tandem Mass Tags) technologies are used for the relative or absolute quantitation of proteins using in vitro isotopic labeling of peptides. The principles and detection methods of both are essentially similar, with the tag reagents comprising three parts: a reporter group, a balance group, and a peptide reactive group. Both use various isotopic reagents as tags to label proteins in different samples, which are then mixed into a single sample for liquid chromatography-mass spectrometry analysis to identify differential proteins.
The number of isotopic labels used differs between iTRAQ and TMT technologies, meaning the number of mixed samples that can be labeled at one time also differs. iTRAQ technology has 4-8 labels, allowing up to 8 samples to be labeled at once. In contrast, TMT offers up to 16 labels, enabling simultaneous analysis of up to 16 different samples.
If both iTRAQ and TMT technologies are used simultaneously for proteomics quantification of the same samples, due to both being isobaric isotopic labels and iTRAQ and TMT labels being similar, it is not possible to differentiate peptides labeled by the two methods in MS2. Ultra-high resolution mass spectrometry is required to meet detection needs, which inevitably leads to more signal interference and may affect quantification accuracy.
Biotech company Biotye Parker uses Thermo Fisher's Q Exactive HF mass spectrometry platform, Orbitrap Fusion mass spectrometry platform, Orbitrap Fusion Lumos mass spectrometry platform combined with Nano-LC, to provideTMT/iTRAQ quantitative proteomics analysisservice. You only need to inform us of the experimental purpose and send the samples, and we will handle all subsequent project matters, including cell culture, cell labeling, protein extraction, protein digestion, peptide separation, mass spectrometry analysis, raw mass spectrometry data analysis, and bioinformatics analysis.
Related services:
Quantitative proteome analysis
Label-based proteomic quantification technologies - iTRAQ, TMT, SILAC
SILAC/Dimethyl labeling-based quantitative proteomics analysis
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