TMTduplex™ Isobaric Mass Tagging Kit

Catalog number: 90063

Thermo Scientific™  Related applications: Protein Mass Spectrometry Analysis

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Description

The Amine-reactive Thermo Scientific TMT™ Isobaric Mass Tagging Kit contains labeling reagents and associated reaction reagents that 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.

Features of the TMTduplex Isobaric Mass Tagging Kit:

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

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.

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.

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.

Related Products
TMTduplex™ Isotopic Label Reagent Set, 5 x 0.8 mg
TMTduplex™ Isobaric Label Reagent Set, 5 x 0.8 mg
TMTzero™ Label Reagent, 5 x 0.8 mg
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Product Size: 15 reactions
Workflow Step: Protein Labeling
Detection Method: Mass Spectrometry
Starting Material: Cell Lysate
For Use With (Equipment): Mass Spectrometer

Contents & storage

Sufficient For: 5 complete duplex (5 x 2-way) experiments
• TMT0 Label Reagent, 5 x 0.8 mg
• TMT2-126 Label Reagent, 5 x 0.8 mg
• TMT2-127 Label Reagent, 5 x 0.8 mg
• Dissolution Buffer (1M triethyl ammonium bicarbonate), 5 mL
• Denaturing Reagent (10% SDS), 1 mL
• Reducing Reagent (0.5M TCEP), 1 mL
• Iodoacetamide, 12 x 9 mg
• Quenching Reagent (50% hydroxylamine), 1 mL
• Trypsin, 2 x 20 µg
• Trypsin Storage Solution, 250 µL
• Albumin, Bovine, 2.5 mg

Store at -20°C

Documents

Manuals & protocols