mTRAQ Reagents

  • The mTRAQ® (Mass differential Tags for Relative and Absolute Quantification) line of quantitative reagents is an offshoot of the iTRAQ® product line.
  • mTRAQ is designed specifically for Multiple Reaction Monitoring (MRM) MS experiments. When researchers have found initial peptides of potential biomarker interest using iTRAQ, they can then use mTRAQ to probe for those specific peptides by MRM.
  • The non-isobaric nature of the mTRAQ analysis allows for maximizing potential differences in the MRM transitions.
  • The mTRAQ reagents can be used to prepare internal standards for all peptides in a sample to be monitored by MRM. This acts as an alternative to synthesizing each individual peptide for MRM.
  • In general, the internal standard is labeled with the Δ8 reagent, while the test samples are labeled with the Δ0 and Δ4 reagents.
  • Peptides labeled with the different mTRAQ labels will share retention time and ionization characteristics. However, different masses will be observed:
(a) Triplex-labeled Lys peptides: 0 Da, 4 Da, or 8 Da difference
(b) Triplex-labeled Arg peptides: 0 Da, 8 Da, or 16 Da difference
  • Related Documents:
Protocol for use of mTRAQ Reagents
Quick Reference Card for use of mTRAQ Reagents
References:
1. DeSouza, L.V., et al., Multiple Reaction Monitoring of mTRAQ-Labeled Peptides Enables Absolute Quantitfication of Endogenous Levels of a Potential Cancer Marker in Cancerous and Normal Endometrial Tissues. J. Proteome Res., 7(8), 3525-3534 (2008). Abstract
2. Kang, U.-B., et al., Quantitative Analysis of mTRAQ-Labeled Proteome Using Full MS Scans. J. Proteome Res., 9(7), 3750-3758 (2010). Abstract
3. Yoon, J.Y., et al., High-throughput peptide quantification using mTRAQ reagent triplex. BMC Bioinformatics, 12(Suppl 1), S46 (2011). Article
4. Kanie, T., et al., Genetic Reevaluation of the Role of F-Box Proteins in Cyclin D1 Degradation. Mol. Cell. Biol., 32(3), 590-605 (2012). Abstract
5. Yin, H-R., et al., Hyperplex-MRM: A Hybrid Multiple Reaction Monitoring Method Using mTRAQ/iTRAQ Labeling for Multiplex Absolute Quantification of Human Colorectal Cancer Biomarker. J. Proteome Res., 12(9), 3912-3919 (2013). Abstract
6. Oppermann, F.S., et al., Comparison of SILAC and mTRAQ Quantification for Phosphoproteomics on a Quadrupole Orbitrap Mass Spectrometer. J. Proteome Res., 12(9), 4089-4100 (2013). Abstract
7. Yeom, J., et al., mTRAQ-based quantitative analysis combined with peptide fractionation based on cysteinyl peptide enrichment. Anal. Biochem., 477, 41-49 (2015). Abstract
8. Dvoráková, M., et al., Transgelin is upregulated in stromal cells of lymph node positive breast cancer. J. Proteomics, 132, 103-111 (2016). Abstract
9. Eidhammer, I., et al., "Targeted Quantification - Selected Reaction Monitoring", from Computational and Statistical Methods for Protein Quantification by Mass Spectrometry. John Wiley & Sons (Chichester, UK), ISBN 978-1-119-96400-1, pp. 218-234 (2013).
10. Lai, X., "Advances in Proteomic Methods", from Bioinformatics of Human Proteomics (ed. Xiangdong Wang). Springer (Dordrecht, Netherlands), ISBN 978-94-007-5810-0, pp. 127-167 (2013).
11. Couto, N., et al., "Making Sense Out of the Proteome: the Utility of iTRAQ and TMT", from New Developments in Mass Spectrometry: Quantitative Proteomics (eds. Claire Eyers & Simon Gaskell). Royal Society of Chemistry (Cambridge, UK), ISBN 978-1-84973-808-8, pp. 51-79 (2014).

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  4352157 Trypsin, TPCK-Treated Trypsin is a serine protease that specifically hydrolyzes peptide bonds at the carboxyl side of Lysine and Arginine residues. This modified trypsin has been treated with N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) to inactivate extraneous chymotryptic activity. This trypsin product also contains calcium chloride, which may improve consistency in digestion. Each package contains 10 vials, with 25 μg in each vial. 
  4370285 Trypsin, TPCK-Treated Trypsin is a serine protease that specifically hydrolyzes peptide bonds at the carboxyl side of lysine and arginine residues. This modified trypsin has been treated with N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) to inactivate extraneous chymotryptic activity. Each package contains 8 vials, with 25 μg in each vial.
  4381664 iTRAQ® Reagent - Multiplex Buffer Kit Contains SDS – Denaturant, TCEP Reducing Reagent, MMTS Cysteine Blocking Reagent, TEAB Buffer and Isopropanol, sufficient material for either two 8-plex Multiplex Kits, three 4-plex Kits, or four mTRAQ® 10 Assay Kits.
  4374771 mTRAQ® Reagent 10 Assay Kit Contains mTRAQ® Reagents Δ0, Δ4, and Δ8, sufficient for 20 mTRAQ assays. One unit of reagent can label up to 100 μg of protein. The kit has 10 tubes each of Δ0, Δ4, and Δ8 reagents, each containing 1 unit of reagent, and the Protocol and Quick Reference Card. Optional Multiplex Buffer kit (Cat. No. 4381664) is to be ordered separately.
  4440015 mTRAQ® Reagent Δ0 50 Unit Pack Contains mTRAQ® Reagents Δ0 for 50 mTRAQ assays. One unit of reagent can label up to 100 μg of protein. Pack has 1 tube each of Δ0 containing 50 units of reagent. Optional Multiplex Buffer kit (Cat. No. 4381664) to be ordered separately.
  4427698 mTRAQ® Reagent Δ4 50 Unit Pack Contains mTRAQ® Reagents Δ4 for 50 mTRAQ assays. One unit of reagent can label up to 100 μg of protein. Pack has 1 tube each of Δ4 containing 50 units of reagent. Optional Multiplex Buffer kit (Cat. No. 4381664) to be ordered separately.
  4427700 mTRAQ® Reagent Δ8 50 Unit Pack Contains mTRAQ® Reagents Δ8 for 50 mTRAQ assays. One unit of reagent can label up to 100 μg of protein. Pack has 1 tube each of Δ8 containing 50 units of reagent. Optional Multiplex Buffer kit (Cat. No. 4381664) to be ordered separately.