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Amine-reactive, Duplex Tandem Mass Tag Reagents


Measure protein expression in two conditions simultaneously.

TMTduplex Tandem Mass Tag Reagents

Amine-reactive Thermo Scientific TMT™ Isobaric Mass Tagging Kits and Reagents enable multiplex quantitation of proteins extracted from cells and tissues using tandem mass spectrometry.

Tandem Mass Tag™ 2-plex (TMTduplex™) Reagents are sets of isobaric compounds (i.e., same mass and structure, also called isotopomers) that are NHS-activated for covalent, irreversible labeling of primary amines (–NH2) groups. Each isobaric reagent contains a different number of heavy isotopes in the mass reporter region, which results in a unique reporter mass during tandem MS/MS for sample identification and relative quantitation. The reagents label all peptides prepared from cell or tissue samples for analysis of two samples in a single MS analysis.

Highlights:

  • Powerful – concurrent MS analysis of two samples increases throughput and enables relative quantitation of two different treatment conditions
  • 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 data sets
  • Robust – consistent chemistry allows efficient transition from method development to multiplex quantitation, enabling biomarker discovery research
  • 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™ 1.0 and above

Applications:

  • Protein expression profiling of normal vs. disease states or control vs. treated
  • Measurement of two 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 quantitation of hundreds to thousands of proteins in a single experiment

Product Details:

Tandem Mass Tag (TMT) Reagents are chemical tags that enable identification and quantitation of proteins in different samples using tandem mass spectrometry. The amine-reactive, NHS-ester-activatedcompounds covalently attach to the peptide amino terminus and free amino termini of lysine residues of peptides and proteins with high efficiency, thereby labeling all peptides in a given sample regardless of enzyme used for digestion. Since TMT reagents share an identical structure and mass (i.e., isotopomers), labeled peptides co-elute during LC separation and are co-isolated during MS/MS analysis, resulting in fewer missing peptide identifications among samples. During MS/MS analysis, each isobaric tag is also fragmented to produce a unique reporter ion mass that is used for sample identification and quantitation. Protein quantitation is accomplished by comparing the relative intensities of the two reporter ions in the MS/MS spectra.

TMT chemical structures for tandem mass tagging of amine groups
Structural design of the amine-reactive Tandem Mass Tag™ Reagents. A. Functional regions of the TMT reagent structure (TMT0) including MS/MS fragmentation sites by higher energy collision dissociation (HCD) and electron transfer dissociation (ETD). B. TMTduplex reagent structures with 13C and 15N heavy isotope positions (red asterisks).

The TMTduplex isobaric reagent set allows protein profiling for two conditions using tandem MS analysis. The TMTzero tag allows testing and optimization of sample preparation, labeling, fractionation and MS fragmentation for peptide identification and reporter detection without using the more costly isotope-labeled compounds. The TMTduplex isotopic reagent set combines the TMTzero tag with one of the TMT6 tags for quantitative protein profiling of two conditions at the MS level similar to SILAC.

Procedure summary for parallel isobaric labeling for tandem mass spectrometry
Procedure summary for MS experiments with Thermo Scientific TMT Isobaric Mass Tagging Reagents. This example uses the TMTsixplex Reagent Set. 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 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. Tandem Mass Tag Reagents combined with the industry-leading Thermo Scientific instruments and software provide integrated total system solutions for quantitative protein expression analysis.

References:

  1. 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.
  2. Dayon, L., et al. (2008). Relative quantification of proteins in human cerebrospinal fluids by MS/MS using 6-plex isobaric tags. Anal. Chem. 80(8):2921-31.
  3. Nilsson, C.L. et al. (2010). Quantitative phosphoproteomic analysis of the STAT3/IL-6/HIF1alpha signaling network: an initial study in GSC11 glioblastoma stem cells. J. Proteome Res. 9:430-43.
  4. Major. M., et al. (2008). Poster: Selective enrichment and quantitation of phosphoproteins involved in cell proliferation. Thermo Fisher Scientific, Rockford, IL 61101 and Cambridge, MA 02139.

Related Resources:

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

Related Products:

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


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
90063 TMTduplex Isobaric Mass Tagging Kit
Formulation: Multi-component kit with TMT0, TMT2-126 and TMT2-127 label reagents
Sufficient For: 5 complete duplex (5 x 2-way) experiments

Kit Contents:
TMT0 Label Reagent, 5 x 0.8mg
TMT2-126 Label Reagent, 5 x 0.8mg
TMT2-127 Label Reagent, 5 x 0.8mg
Dissolution Buffer (1M triethyl ammonium bicarbonate), 5mL
Denaturing Reagent (10% SDS), 1mL
Reducing Reagent (0.5M TCEP), 1mL
Iodoacetamide, 12 x 9mg
Quenching Reagent (50% hydroxylamine), 1mL
Trypsin, 2 x 20μg
Trypsin Storage Solution, 250μL
Albumin, Bovine, 2.5mg
15-rxn kit Product Instructions for product #90063 TMTduplex Isobaric Mass Tagging Kit MSDS for product #90063 TMTduplex Isobaric Mass Tagging Kit Certificate of Analysis for product #90063 TMTduplex Isobaric Mass Tagging Kit $728.00

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90065 TMTduplex Isobaric Label Reagent Set, 5 x 0.8mg
Formulation: Set of TMT2-126, TMT2-127 label reagents
Sufficient For: 5 duplex (5 x 2-way) experiments

Kit Contents:
TMT2-126 Label Reagent, 5 x 0.8mg
TMT2-127 Label Reagent, 5 x 0.8mg
10-rxn set Product Instructions for product #90065 TMTduplex Isobaric Label Reagent Set, 5 x 0.8mg MSDS for product #90065 TMTduplex Isobaric Label Reagent Set, 5 x 0.8mg Certificate of Analysis for product #90065 TMTduplex Isobaric Label Reagent Set, 5 x 0.8mg $560.00

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90060 TMTduplex Isotopic Label Reagent Set, 5 x 0.8mg
Formulation: Set of TMT0 and TMT6-127 label reagents
Sufficient For: 5 duplex (5 x 2-way) experiments

Kit Contents:
TMT0 Label Reagent, 5 x 0.8mg
TMT6-127 Label Reagent, 5 x 0.8mg
10-rxn set Product Instructions for product #90060 TMTduplex Isotopic Label Reagent Set, 5 x 0.8mg MSDS for product #90060 TMTduplex Isotopic Label Reagent Set, 5 x 0.8mg Certificate of Analysis for product #90060 TMTduplex Isotopic Label Reagent Set, 5 x 0.8mg $538.00

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90067 TMTzero Label Reagent, 5 x 0.8mg
Formulation: TMT0-126 label reagent
Sufficient For: Controls in 5 experiments

Kit Contents:
TMT0 Label Reagent, 5 x 0.8mg
5-rxn set Product Instructions for product #90067 TMTzero Label Reagent, 5 x 0.8mg MSDS for product #90067 TMTzero Label Reagent, 5 x 0.8mg Certificate of Analysis for product #90067 TMTzero Label Reagent, 5 x 0.8mg $180.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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