For protein expression analysis of sulfhydryl components by mass spectrometry.
The cysteine-reactive Thermo Scientific iodoTMT* (Tandem Mass Tag*) Reagents and Labeling Kit enable measurement of protein and peptide cysteine modifications (S-nitrosylation, oxidation and disulfide bridges) by multiplex quantitative mass spectrometry.
Tandem Mass Tag* (TMT*) Reagents enable concurrent identification and multiplexed quantitation of proteins in different samples using tandem mass spectrometry. The iodoTMT* Reagents are sets of isobaric (mass and structure) isomers that are iodoacetyl-activated for covalent, irreversible labeling of sulfhydryl (–SH) groups. Similar to iodoacetamide, iodoTMT Reagents react specifically with reduced cysteines (Cys) in peptides and proteins. IodoTMT Reagents can be differentiated by mass spectrometry (MS), enabling quantitation of the relative abundance of cysteine modifications, such as S-nitrosylation, oxidation and disulfide bonds, in cultured cells grown or treated with different conditions.
Specific – only reacts with sulfhydryl groups (reduced, unmodified cysteine residues)
Irreversible – labeled proteins and peptides are not susceptible to reducing agents
Flexible – options for duplex isotopic (MS) or sixplex isobaric (MS/MS) quantitation
Versatile – can be used to study cysteine modifications (e.g. di-sulfide bridges, S-nitrosylation)
IodoTMT Products replace our previously offered CysTMT* Reagents, which utilized a dithiopyridine reactive group to reversibly label cysteine sulfhydryls. For information about customized reagent sets, please contact our Large Volume and Custom Sales Department.
Thermo Scientific iodoTMT Reagents.(A) iodoTMTzero reagent showing key structural features, as well as CID (collision-induced dissociation) and ETD (electron-transfer dissociation) sites. (B) Structures of iodoTMTsixplex reagents showing the mass (Da) of each reporter and the distribution of normalizing isotopes. Learn about the iodoacetyl reaction scheme.
Workflow schematic for sixplex reagent experiment.(A) Six different sample conditions can be prepared for labeling with Thermo Scientific iodoTMTsixplex Reagents. Labeled proteins are combined before iodoTMT peptide enrichment using immobilized anti-TMT antibody resin. (B) Subsequent LC-MS/MS analysis of isobaric reporter ions provides quantitation of test samples.
Thompson, A., et al. (2003). Tandem mass tags: a novel quantification strategy for comparative analysis of complex protein mixtures by MS/MS. Anal. Chem. 75:1895-204.
Forrester MT, et al. (2009). Detection of protein S-nitroylsation with the biotin technique. Free Radic Biol Med. 46(2):119-126.
Gygi, S.P., et al. (1999). Quantitative analysis of complex protein mixtures using isotopecoded affinity tags. Nat. Biotech. 17:994-9.
Murray CI, et al. (2012) Identification and quantification of S-nitrosylation by cysteine reactive tandem mass tag switch assay. Mol Cell Proteomics. 11(2): M111.013441.