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™ 6-plex (TMTsixplex™) 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 up to six samples in a single MS analysis.

Features of this TMTsixplex Label Reagent Set:

• Powerful – concurrent MS analysis of multiple samples increases sample throughput and enables relative quantitation of up to six 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 – 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 identification and quantitation from multiple samples of cells, tissue or biological fluids
• Protein expression profiling of normal vs. disease states or control vs. treated
• Measurement of up to six 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

Changes in protein expression and post-translational modifications are essential mechanisms of biological regulation and disease. Advancements in liquid chromatography (LC), mass spectrometry (MS) instrumentation and bioinformatics now enable researchers to identify thousands of proteins in a given sample with a high degree of confidence.  However, reproducible detection and quantitation of different proteins within samples using mass spectrometry is challenging due to variability in peptide separation, ionization and detection.

Tandem Mass Tag (TMT) Reagents are isobaric chemical tags that enable concurrent identification and quantitation of proteins in different samples using tandem mass spectrometry. The amine-reactive, NHS-ester-activated compounds 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 six reporter ions in the MS/MS spectra.

The Tandem Mass Tag (TMT) Reagent family consists of TMTzero, TMTduplex, TMTsixplex and TMT10plex sets which are specially designed to enable a rapid and cost-effective transition from method development to high-throughput protein quantitation. 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 TMTsixplex reagent set allows sixplex protein profiling for multiple conditions, including time courses, dose responses, replicates or multiple sample comparisons. Each TMT tag is based on the same chemical structure, eliminating the need to modify labeling conditions or HPLC separation conditions between experiments.

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.

More Product Data
• Quantitative cancer stem cell phosphoprotein profiling

Related Products
TMTsixplex™ Isobaric Mass Tagging Kit

For high resolution analysis of the TMT-labeled peptides, the recommended LC column for Nanospray Flex source is Acclaim PepMap 100 C18 LC Column (Cat. No. 164942 or 164939). For EASY-Spray source, the recommended LC column is EASY-Spray C18 LC Column (Cat. No. ES802 or ES803).