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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. Amine-reactive Thermo Scientific TMT Isobaric Mass Tagging Kits and Reagents enable multiplex quantitation of proteins extracted from cells and tissues from up to 6 samples using tandem mass spectrometry.
Features of TMTsixplex Label Reagents and Kits:
• Powerful—multiplexed MS analysis increases sample throughput and enables relative quantitation of up to six different samples derived from cells, tissues or biological fluids
• Robust—consistent chemistry allows efficient transition from method development to multiplex quantitation, enabling biomarker discovery research
• Efficient—amine-reactive NHS-ester activated reagents ensures greater than 95% labeling of all peptides regardless of protein sequence or proteolytic enzyme specificity
• Compatible—optimized for use with high resolution Thermo Scientific Orbitrap MS/MS platforms, with data analysis fully supported by Proteome Discoverer™ 2.3 and above
Applications of TMTsixplex Label Reagents and Kits:
• 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
• 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
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.
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 high resolution Thermo Scientific Orbitrap instruments and Proteome Discoverer software provide integrated total system solutions for quantitative protein expression analysis. 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. ES902 or ES903).