Amine-reactive Thermo Scientific TMT10plex™ Isobaric Mass Tag Labeling Kits and Reagents enable multiplexed protein identification and quantitative analysis by tandem mass spectrometry (MS).
This Tandem Mass Tag™ (TMT™) 10-plex reagent set contains ten different isobaric compounds with the same mass and chemical structure (i.e., isotopomeric) composed of an amine-reactive NHS-ester group, a spacer arm and a mass reporter. The reagent set enables up to ten different peptide samples prepared from cells or tissues to be labeled in parallel and then combined for analysis. For each sample, a unique reporter mass (i.e., TMT10 126-131Da) in the low-mass region of the high-resolution MS/MS spectrum is used to measure relative protein expression levels during peptide fragmentation and tandem mass spectrometry.
- Powerful – concurrent MS analysis of multiple samples increases sample throughput and enables relative quantitation of up to ten different samples derived from cells, tissues or biological fluids
- Consistent – identical reagent structure and performance among TMTzero™, TMTduplex™, TMTsixplex™ and TMT10plex™ reagents allows efficient transition from method development to multiplex quantitation, enabling biomarker discovery research across platforms and datasets
- Robust – increased multiplex capability results in fewer missing quantitative values among samples and higher confidence among replicates
- Efficient – amine-reactive, NHS-ester-activated reagents ensure efficient labeling of all peptides regardless of protein sequence or proteolytic enzyme specificity
- Compatible – optimized for use with high resolution Thermo Scientific MS/MS platforms, such as the Q Exactive, Orbitrap Elite™ and Orbitrap Fusion™ Tribrid™ instruments with data analysis fully supported by Proteome Discoverer™ 1.4
- Protein identification and quantitation from multiple samples of cells, tissue or biological fluids
- Protein expression profiling of normal vs. disease states or control vs. treated
- Multiplex up to ten different samples concurrently in a single experiment
- Quantitative analysis of proteins for which no antibodies are available
- Identification and quantitation of membrane and post-translationally modified proteins
- Identification and quantification of hundreds to thousands of proteins in a single experiment
The Tandem Mass Tag (TMT) Reagents are specially designed to enable identification and quantitation of proteins in different samples using tandem mass spectrometry. The TMT10plex Label Reagents share an identical structure with TMTzero, TMTduplex, and TMTsixplex Reagents but contain different numbers and combinations of 13C and 15N isotopes in the mass reporter. The different isotopes result in a 10-plex set of tags that have monoisotopic mass differences in the reporter that can be detected using high resolution Orbitrap MS instruments.
|Structural design of the amine-reactive Tandem Mass Tag™ Reagents.
A. Functional regions of the reagent structure including MS/MS fragmentation sites by higher energy collision dissociation (HCD) and electron transfer dissociation (ETD). B. TMT10plex reagent structures with 13C and 15N heavy isotope positions (red asterisks).
Advantages of the TMT10plex Label Reagents include increased multiplex relative quantitation, increased sample throughput, and fewer missing quantitative values among samples. TMT10plex Label Reagents are ideal for analysis of multiple protein samples from inhibitor dose response experiments, time course experiments or biological replicates.
|Procedure summary for MS experiments with Thermo Scientific TMT Isobaric Mass Tagging Reagents. Protein extracts isolated from cells or tissues are reduced, alkylated and digested overnight. Samples are labeled with the TMT Reagents and then mixed before sample fractionation and clean up. Labeled samples are analyzed by high resolution Orbitrap LC-MS/MS before data analysis to identify peptides and quantify reporter ion relative abundance.
TMT Reagents are provided as standalone sets or in optimized kit formats containing all necessary reagents and controls for maximum flexibility, convenience and reliability. When combined with the industry-leading, high resolution Thermo Scientific Orbitrap instruments and software, TMT10plex Reagents provide integrated total solutions for quantitative protein expression analysis.
|Example of 10-plex relative quantitation using Thermo Scientific TMT10plex Reagents. BSA tryptic digests labeled with TMT10plex reagents (TMT10 126-131Da ) were mixed 16:8:4:2:1:1:2:4:8:16 and analyzed by high resolution (Oribitrap LC-MS). The relative abundance of the target protein or peptide fragment in ten different samples is easily measured by comparing the reporter ions generated by MS/MS fragmentation of the different mass tags.
- Thompson, A., et al. (2003). Tandem mass tags: a novel quantification strategy for comparative analysis of complex protein mixtures by MS/MS. Anal. Chem. 75(8): 1895-1904.
- McAllister, G.C., et al. (2012). Increasing the multiplexing capacity of TMTs using reporter ion isotopologues with isobaric masses. Anal. Chem. 84(17): 7469-78.
- Werner, T., et al (2012). High-resolution enabled TMT 8‑plexing. Anal Chem. 2012. 84(16):7188-94.
Mass Spec Sample Prep Workflow (selection guide)
Mass spectrometry technical guide
Quantitative proteomics technical guide
Service providers that use our mass tagging products
TMT Technology - opens Proteome Sciences website
Other TMT Reagents
SILAC Protein Quantitation Kits
Ubiquitin, Glycoprotein and Phosphoprotein Enrichment Kits
Fe-NTA, TiO2, C-18 and Graphite peptide enrichment and clean-up kits
Protease and Phosphatase Inhibitor Cocktails and Tablets
Currently available by special order only.
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815-968-0747 ext. 300 or +800-874-3723 ext. 300