Kit containing complete set of TMT 10-plex Isobaric Label Reagents

Tandem mass tags (TMT) are robust and efficient systems for multiplex, relative protein quantitation of up to 18 samples, derived from cells, tissues, or biological fluids. All mass tagging reagents within a label reagent set have the same nominal mass (i.e., they are isobaric) and chemical structure, composed of a reactive group, a spacer arm (mass normalizer), and a mass reporter. The isobaric nature of the TMT labeling reagents allow for concurrent identification and quantification of proteins in different samples using tandem mass spectrometry (MS).

A quick comparison of TMT vs. TMTpro label reagents is shown in Table 1.

See TMT selection guide

Table 1. Comparison of features of TMT and TMTpro reagents

FeaturesTMTTMTpro
ReactivityAmine-reactive
Cysteine-reactive
Carbonyl-reactive
Amine-reactive
Multiplex2- to 11-plex16- to 18-plex
Isotopes15N1, 13C515N2, 13C
Chemical structure
  • Spacer arm
  • a peptide-reactive labeling group
  • Dimethyl piperidine mass reporter
  • Longer spacer arm
  • a peptide-reactive labeling group
  • Isobutyl proline mass reporter
Reporter ion masses126–131C126–134N
Product formatLabel reagents and KitsLabel reagents
Applications
  • Protein identification
  • Protein quantitation
  • Protein expression profiling

 


Selection guide for TMT reagents

The TMTpro label reagents have increased multiplex capability and can be used for concurrent protein mass spectrometry analysis of 16 to 18 biological samples.

Learn more about TMTpro reagents

PlexReactive chemistry*ProductUnit sizeCat. No.
16-plexAmine-reactiveTMTpro 16-plex Label Reagent Set1 x 5 mg (per tag)A44520
1 x 0.5 mg (per tag)A44521
6 x 0.5 mg (per tag)A44522
TMTpro 16plex Label Reagents in stabilized solution, 96-well plate

6 x 50 µg (per tag)

A58334
18-plexAmine-reactiveTMTpro 18-plex Label Reagent Set1 x 5 mgA52045
6 x 0.5 mgA52047
TMTpro 18plex Label Reagents in stabilized solution, 96-well plate4 x 50 µg (per tag)A58335
18-plexAmine-reactiveTMTpro-134C and
TMTpro-135N Label Reagents
1 x 5 mgA52046
6 x 0.5 mgA52048

*All proteins and peptides.

Additional TMTpro reagents

Reagent typeReactive chemistry*ProductUnit sizeCat. No.
ControlAmine-reactiveTMTpro Zero Label Reagent1 x 5 mgA44518
5 x 0.5 mgA44519
Specialized for 2D multiplexingAmine-reactiveSuper Heavy TMTpro Label Reagent2 mgA52040

*All proteins and peptides.

The TMT reagents and kits are categorized based on reactive chemistry groups. See all amine reactive 2- to 11-plex reagents below.

Learn more about TMT reagents

MultiplexReactive chemistry*ProductUnit sizeCat. No.
2-plexAmine reactiveTMTduplex Isobaric Mass Tagging Kit15 reactions90063
2-plexAmine reactiveTMTduplex Isobaric Label Reagent Set5 x 0.8 mg90065
2-plexAmine reactiveTMTduplex Isotopic Label Reagent Set5 x 0.8 mg90060
6-plexAmine reactiveTMTsixplex Isobaric Label Reagent Kit 35 reactions90064
Contact us90064B
6-plexAmine reactiveTMTsixplex Isobaric Label Reagent Set1 x 0.8 mg (per tag)90061
2 x 0.8 mg (per tag)90062
5 x 0.8 mg (per tag)90066
2 x 5 mg (per tag)90068
10-plexAmine reactiveTMT10plex Isobaric Mass Tagging Kit30 reactions90113
10-plexAmine reactiveTMT10plex Isobaric Label Reagent Set1 x 0.8 mg (per tag)90110
3 x 0.8 mg (per tag)90111
1 x 5 mg (per tag)90406
8 x 0.2 mg (per tag)90309
TMT 10plex Label Reagents in stabilized solution, 96-well plate8 x 40 µg (per tag)A58332
11-plexAmine reactiveTMT10plex Isobaric Label Reagent Set plus TMT11-131C Label Reagent1 x 5 mg (per tag)A34808
3 x 0.8 mg (per tag)A37725
TMT 11plex Label Reagents in stabilized solution, 96-well plate8 x 40 µg (per tag)A58333
11-plexAmine reactiveTMT11-131C Label Reagent1 x 5 mg (per tag)A34807
3 x 0.8 mg (per tag)A37724

*All proteins and peptides.

Additional TMT reagents

Reagent typeReactive chemistry*ProductUnit sizeCat. No.
ControlAmine-reactiveTMT Zero Label Reagent5 x 0.8 mg90067
Specialized for 2D multiplexingAmine-reactiveSuper Heavy TMT Label Reagent2 mgA43073

*All proteins and peptides.

The TMT reagents and kits are categorized based on reactive chemistry groups. See all cysteine-reactive and carbonyl-reactive 6plex reagents below.

Learn more about TMT reagents

MultiplexReactive chemistry*ProductUnit sizeCat. No.
6-plexCysteine-reactive*iodoTMTsixplex Isobaric Mass Tagging Kit30 reactions90103
6-plexCysteine-reactive*iodoTMTsixplex Isobaric Label Reagent Set1 x 0.2 mg (per tag)90101
5 x 0.2 mg (per tag)90102
6-plexCarbonyl-reactive**aminoxyTMTsixplex Label Reagent Set1 x 0.2 mg (per tag)90401
5 x 0.2 mg (per tag)90402

* Peptides with reduced cysteine (Cys)
** Peptides with carbonyl

Control reagent required for Cysteine-reactive proteins

Reagent typeReactive chemistry*ProductUnit sizeCat. No.
ControlCysteine-reactiveiodoTMTzero Label Reagent5 x 0.2 mg90100

*Peptides with reduced cysteine (Cys)

The AccelerOme TMT labeling kits enable multiplex relative quantitation by protein mass spectrometry. These kits are available in combo packs containing a sample prep module, a TMT module, and a peptide retention time calibration mixture.

Learn more about AccelerOme reagents

MultiplexReactive chemistry*ProductUnit sizeCat. No.
11-plexAmine-reactiveAccelerOme TMT11plex MS Sample Preparation Kit, 10–30 µg3 x 11 reactionsA50946
11-plexAmine-reactiveAccelerOme TMT11plex MS Sample Preparation Kit, 30–100 µg1 x 11 reactionsA50947
3 x 11 reactionsA50948
16-plexAmine-reactiveAccelerOme TMTpro 16plex MS Sample Preparation Kit, 10–30 µg2 x 16 reactionsA50949
16-plexAmine-reactiveAccelerOme TMTpro 16plex MS Sample Preparation Kit, 30–100 µg1 x 16 reactionsA50950
2 x 16 reactionsA50951

*All proteins and peptides

The TMTpro label reagents have increased multiplex capability and can be used for concurrent protein mass spectrometry analysis of 16 to 18 biological samples.

Learn more about TMTpro reagents

PlexReactive chemistry*ProductUnit sizeCat. No.
16-plexAmine-reactiveTMTpro 16-plex Label Reagent Set1 x 5 mg (per tag)A44520
1 x 0.5 mg (per tag)A44521
6 x 0.5 mg (per tag)A44522
TMTpro 16plex Label Reagents in stabilized solution, 96-well plate

6 x 50 µg (per tag)

A58334
18-plexAmine-reactiveTMTpro 18-plex Label Reagent Set1 x 5 mgA52045
6 x 0.5 mgA52047
TMTpro 18plex Label Reagents in stabilized solution, 96-well plate4 x 50 µg (per tag)A58335
18-plexAmine-reactiveTMTpro-134C and
TMTpro-135N Label Reagents
1 x 5 mgA52046
6 x 0.5 mgA52048

*All proteins and peptides.

Additional TMTpro reagents

Reagent typeReactive chemistry*ProductUnit sizeCat. No.
ControlAmine-reactiveTMTpro Zero Label Reagent1 x 5 mgA44518
5 x 0.5 mgA44519
Specialized for 2D multiplexingAmine-reactiveSuper Heavy TMTpro Label Reagent2 mgA52040

*All proteins and peptides.

The TMT reagents and kits are categorized based on reactive chemistry groups. See all amine reactive 2- to 11-plex reagents below.

Learn more about TMT reagents

MultiplexReactive chemistry*ProductUnit sizeCat. No.
2-plexAmine reactiveTMTduplex Isobaric Mass Tagging Kit15 reactions90063
2-plexAmine reactiveTMTduplex Isobaric Label Reagent Set5 x 0.8 mg90065
2-plexAmine reactiveTMTduplex Isotopic Label Reagent Set5 x 0.8 mg90060
6-plexAmine reactiveTMTsixplex Isobaric Label Reagent Kit 35 reactions90064
Contact us90064B
6-plexAmine reactiveTMTsixplex Isobaric Label Reagent Set1 x 0.8 mg (per tag)90061
2 x 0.8 mg (per tag)90062
5 x 0.8 mg (per tag)90066
2 x 5 mg (per tag)90068
10-plexAmine reactiveTMT10plex Isobaric Mass Tagging Kit30 reactions90113
10-plexAmine reactiveTMT10plex Isobaric Label Reagent Set1 x 0.8 mg (per tag)90110
3 x 0.8 mg (per tag)90111
1 x 5 mg (per tag)90406
8 x 0.2 mg (per tag)90309
TMT 10plex Label Reagents in stabilized solution, 96-well plate8 x 40 µg (per tag)A58332
11-plexAmine reactiveTMT10plex Isobaric Label Reagent Set plus TMT11-131C Label Reagent1 x 5 mg (per tag)A34808
3 x 0.8 mg (per tag)A37725
TMT 11plex Label Reagents in stabilized solution, 96-well plate8 x 40 µg (per tag)A58333
11-plexAmine reactiveTMT11-131C Label Reagent1 x 5 mg (per tag)A34807
3 x 0.8 mg (per tag)A37724

*All proteins and peptides.

Additional TMT reagents

Reagent typeReactive chemistry*ProductUnit sizeCat. No.
ControlAmine-reactiveTMT Zero Label Reagent5 x 0.8 mg90067
Specialized for 2D multiplexingAmine-reactiveSuper Heavy TMT Label Reagent2 mgA43073

*All proteins and peptides.

The TMT reagents and kits are categorized based on reactive chemistry groups. See all cysteine-reactive and carbonyl-reactive 6plex reagents below.

Learn more about TMT reagents

MultiplexReactive chemistry*ProductUnit sizeCat. No.
6-plexCysteine-reactive*iodoTMTsixplex Isobaric Mass Tagging Kit30 reactions90103
6-plexCysteine-reactive*iodoTMTsixplex Isobaric Label Reagent Set1 x 0.2 mg (per tag)90101
5 x 0.2 mg (per tag)90102
6-plexCarbonyl-reactive**aminoxyTMTsixplex Label Reagent Set1 x 0.2 mg (per tag)90401
5 x 0.2 mg (per tag)90402

* Peptides with reduced cysteine (Cys)
** Peptides with carbonyl

Control reagent required for Cysteine-reactive proteins

Reagent typeReactive chemistry*ProductUnit sizeCat. No.
ControlCysteine-reactiveiodoTMTzero Label Reagent5 x 0.2 mg90100

*Peptides with reduced cysteine (Cys)

The AccelerOme TMT labeling kits enable multiplex relative quantitation by protein mass spectrometry. These kits are available in combo packs containing a sample prep module, a TMT module, and a peptide retention time calibration mixture.

Learn more about AccelerOme reagents

MultiplexReactive chemistry*ProductUnit sizeCat. No.
11-plexAmine-reactiveAccelerOme TMT11plex MS Sample Preparation Kit, 10–30 µg3 x 11 reactionsA50946
11-plexAmine-reactiveAccelerOme TMT11plex MS Sample Preparation Kit, 30–100 µg1 x 11 reactionsA50947
3 x 11 reactionsA50948
16-plexAmine-reactiveAccelerOme TMTpro 16plex MS Sample Preparation Kit, 10–30 µg2 x 16 reactionsA50949
16-plexAmine-reactiveAccelerOme TMTpro 16plex MS Sample Preparation Kit, 30–100 µg1 x 16 reactionsA50950
2 x 16 reactionsA50951

*All proteins and peptides

TMTpro reagents overview

The TMTpro 16-plex and 18-plex label reagents are the next generation of TMT reagents (TMT tags) that offer increased multiplex capability and sample throughput [1]. Concurrent protein mass spectrometry analysis of up to 18 samples, derived from cells, tissues, or biological fluids can be performed with accuracy and precision.

The amine-reactive TMTpro reagents are used for labeling tryptic peptides irrespective of their protein sequence or enzyme specificity. The isobaric TMTpro label reagents are similar in design to the original TMT label reagents except that they have a longer spacer region and isobutyl proline mass reporter region (Figure 1). After MS/MS fragmentation, each TMTpro tag generates a unique reporter mass (126–134 Da) in the low-mass region of the high-resolution MS/MS spectrum that is used for relative protein quantitation.

The workflow for TMTpro and original TMT label reagents is identical (Figure 2). Additional mass spectrometry reagents such as standards, buffers, antibody and sample prep kit can be purchased separately (Table 2).

TMTpro zero label reagent (serves as a ‘light’ tag) is used for method development and labelling optimization before multiplexed analysis of labeled samples. It is a suitable alternative to costly isotope-labeled compounds and allows testing and optimization of sample preparation, labeling, fractionation, and fragmentation, for peptide identification, and reporter detection.

Functional regions of TMTPro reagents’ chemical structure

Figure 1. Structural design of TMTpro label reagents. The functional regions of the TMTpro reagents’ structure including MS/MS fragmentation site by higher-energy collision dissociation (HCD).

Schematic representation of TMT and TMTPro reagents’ workflow

Figure 2. Schematic summary for MS experiments using TMTpro or TMT original isobaric mass tagging reagents. Protein extracts isolated from cells or tissues are reduced, alkylated, and then digested using the EasyPep Mini MS Sample Prep kit or an equivalent method [2]. Samples are then labeled with the TMT or TMTpro reagents before sample mixing, fractionation, and cleanup. Labeled samples are analyzed on a high-resolution Orbitrap LC-MS/MS mass spectrometer before data analysis to identify peptides and quantify relative abundance of reporter ions.

Table 2. Additional MS reagents.

Workflow stepProductUnit sizeCat. No.
TMT standardPierce TMT11plex Yeast Digest Standard20 µgA40938
5 x 20 µgA40939
TMT labeling1M Triethylammonium bicarbonate (TEAB) for TMT experiments50 mL90114
TMT quench50% Hydroxylamine for TMT experiments5 mL90115
Enrichment of TMT-labeled peptidesTMT Monoclonal Antibody (25D5)100 µg90075
Enrichment of TMT-labeled peptidesImmobilized Anti-TMT Antibody Resin6 mL90076
Enrichment of TMT-labeled peptidesTMT Elution Buffer20 mL90104
TMT-compatible peptide sample prepEasyPep MS Sample Prep Kit8 reactionsA45734
20 reactionsA40006
96 reactionsA45733
TMT-compatible peptide sample prepEasyPep Lysis Buffer100 mLA45735

TMT reagents overview

TMT label reagents (TMT tags) enable multiplex (2-plex to 11-plex) relative quantitation of biological samples using protein mass spectrometry. The chemical structure of TMT reagents consists of a peptide-reactive labeling group, a mass reporter, and a spacer arm (mass normalizer) (Figure 3). Based on the reactive chemistry, the TMT reagents are classified into three categories—amine reactive, cysteine-reactive, and carbonyl-reactive.

  • The amine-reactive reagents are explained in the TMTpro section above. The structure is shown in Figure 3(A).
  • The iodoTMT reagents react specifically with reduced cysteines (Cys) in peptides and proteins, and irreversibly label free sulfhydryl groups (Figure 3(B)).
  • The aminoxyTMT reagents are optimized for efficient labeling and recovery of glycans and other carbonyl-reactive proteins, and offer greater stability of the labeled product (Figure 3(C)).

The TMT 10-plex and 11-plex 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.

Functional regions of the TMT reagents’ chemical structure

Figure 3. Structural design of TMT reagents. The functional regions of the TMT reagents’ structure including MS/MS fragmentation site by higher-energy collision dissociation (HCD) and electron transfer dissociation (ETD) and CID (collision induced dissociation). Amine-reactive, cysteine-reactive, and carbonyl-reactive TMT reagents are shown in (A), (B), and (C).


Liquid, 96-well format overview

TMT labeling reagents in lyophilized format are extremely reactive to moisture and great handling care must be taken to ensure proper functionality. A subset of TMT 10- and 11-plex and TMTpro 16- and 18-plex label reagents have been developed to be stable in liquid format and are pre-aliquoted in 96-well plates for simplified processing.

Highlights include:

  • Support of higher throughput and automation with barcodes
  • Economical with flexible partial plate utility; multiple freeze-thaw stable
  • Simplified processing; no need to reformat lyophilized reagent into multiwell plates
  • Equivalent performance to lyophilized TMT and TMTpro labeling reagents
  • Less starting material required, with labeling ability of 1–10 ug of digested protein
TMTpro 16plex Label Reagents in stablized solution, 96-well plate

TMT labeling reagent in 96-well plate

Bottom view of TMT 96-well plate
(A) Liquid stable TMT/TMTpro
Cut and peel foil for partial TMT plate use
(B) Cut and peel foil seal
Punctured foil seal
(C) Puncture foil seal

Figure 4. TMT labeling reagent in 96-well plate. Flexible format of 96-well TMT labeling reagent in stable solution with barcoding for optional automation use, with an easy to peel or puncture foil seal. For partial plate use, TMT and TMTpro labeling reagents can be thawed and refrozen multiple times with continued high performance.  (A) TMT and TMTpro in stabilized solution pre-aliquoted in 96-well plate. (B) Foil seal can be peeled and cut for partial plate use. (C) Foil seal can easily be punctured permitting partial plate use.

Super Heavy TMT reagents

The Super heavy TMT (shTMT) and shTMTpro label reagents are non-isobaric amine-reactive TMT variants and are a suitable alternative for precursor-based protein quantification [3, 4]. These TMT derivatives contain twice the number of heavy isotopes (+11 mass units) that are dispersed throughout the reporter and spacer regions (Figure 4). They are used as a mass offset for triggering peptide selection. The workflow for Super heavy TMT (shTMT) and shTMTpro label reagents is identical (Figure 5). The shTMT label reagent is used in combination with TMT- or TMTpro-labeled samples and the mass offset from the super heavy-labeled synthetic peptide serves as a trigger for the selection of the targeted peptide in the multiplexed sample. This triggered selection reduces interference/co-isolation to enable more robust quantitation.

Structural design of Super heavy TMTpro label reagent

Figure 5. Chemical structure of Super Heavy TMTpro (shTMTpro) label reagents.

Schematic workflow of Super heavy TMTpro reagents

Figure 6. Super Heavy TMTpro workflow.

AccelerOme reagents

The AccelerOme reagents are the newest additions to the next generation tandem mass tags (TMT) to support automated sample preparation and TMT labeling on the AccelerOme Automated Sample Preparation Platform. The integrated instrument and reagent platform enables automated sample handling of up to 36 samples for the reduction, alkylation, and digestion of proteins, followed by TMT or TMTpro labeling and clean-up. The TMT or TMTpro-labeled samples are fit-for-purpose for analysis using high resolution Thermo Scientific Orbitrap LC-MS/MS instruments (Figure 6).

Schematic representation of AccelerOme automated sample preparation system

Figure 7. Supporting kits and workflow of AccelerOme automated sample preparation system.

Videos and webinars

TMT10plex reagents in action

Learn how to prepare and label peptide samples with tandem mass tags for quantitative proteomics analysis.

Resources
accelerome-brochure-400x350

AccelerOme Automated Sample Preparation Platform brochure

 Download

proteome-discoverer-software-application-note-400x350-2

Proteome Discoverer 2.3 software application note

 Download

References
Stylesheet for Classic Wide Template adjustments

For Research Use Only. Not for use in diagnostic procedures.