Principles of TMT and iTRAQ Technologies
iTRAQ (Isobaric Tag for Relative Absolute Quantitation) and TMT (Tandem Mass Tags) are peptide in vitro labeling quantitative technologies developed by AB Sciex and Thermo, respectively. These technologies are based on in vitro isobaric labeling for relative or absolute quantification. They use various isobaric isotopic reagents as tags to simultaneously label different protein samples, which are then mixed together for detection.
These isobaric isotopic tag reagents consist of three different chemical tags: the reporter part, the peptide reactive part, and the balance part. The reporter part is selected based on the experimental design to determine the number of labels, forming 2-18 types of isobaric isotopic tags. The peptide reactive part connects the iTRAQ/TMT tags with the peptide N-terminus and lysine side chain residues for labeling digested peptides. The balance part ensures that the same peptide segments in different samples labeled by iTRAQ/TMT reagents have the same mass-to-charge ratio (m/z). In tandem mass spectrometry (MS/MS), the same peptide segments labeled with different iTRAQ/TMT reagents have identical molecular weights detected in the first stage mass spectrometry (MS1), making it impossible to distinguish labeled peptides. However, in the second stage mass spectrometry (MS2) detection, the balance group of the iTRAQ/TMT label detaches, producing signal ions with different m/z peaks. Protein quantification can be performed based on the height and area of these peaks in the second stage mass spectrometry.
Biotech company Biotech Park uses Thermo Fisher's Q Exactive HF mass spectrometry platform, Orbitrap Fusion mass spectrometry platform, and Orbitrap Fusion Lumos mass spectrometry platform combined with Nano-LC to provideTMT/iTRAQ quantitative proteomics analysisservices. You only need to inform us of the experimental objectives and send the samples. 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 proteomics analysis
Tag-based protein quantification technologies - iTRAQ, TMT, SILAC
SILAC/Dimethyl labeling-based quantitative proteomics analysis
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