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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 amine groups (—NH2). The TMTduplex isobaric reagent set allows protein profiling for two conditions using tandem MS analysis.
Features of TMTduplex isobaric and isotopic label reagent sets:
• Powerful—concurrent MS analysis of two samples increases throughput and enables relative quantitation of two different treatment conditions (up to five experiments per set)
• 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
• 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 Orbitrap MS/MS platforms, with data analysis fully supported by Proteome Discoverer 2.4 and above
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. TMTduplex isobaric label reagents comprise “heavy” TMT2-126 and TMT2-127 label reagents for multiplexing. Both sets of label reagents have the same structure. 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 mass spectrometry analysis. During MS/MS analysis, each TMT 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. Our TMTduplex reagents label all peptides for analysis of two samples in a single MS analysis.
• 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
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 instruments and software provide integrated total system solutions for quantitative protein expression analysis.