Thermo Scientific™ aminoxyTMTsixplex™ Label Reagent Set

Description

• The aminoxyTMTsixplex Label Reagents share an identical structure but contain different numbers of 13C and 15N isotopes in the mass reporter
• The optimized labeling procedure can be completed in one hour
• After tag labeling, samples are quenched and cleaned-up using HILIC solid-phase extraction before mass spectrometry analysis
• The six compounds of the aminoxyTMTsixplex™ Reagent Set have the same mass (i.e., isobaric) and chemical structure (carbonyl-reactive aminoxy group, spacer arm, and mass reporter)
• However, the specific distribution of 13C and 15N isotopes on either side of the HCD or ETD MS/MS fragmentation site in each reagent results in a unique reporter mass (126-131Da) in the low mass region
• This set of reporter ions is used to measure the relative abundance of labeled molecules in a combined (multiplexed) MS sample representing six different treatment conditions
• For glycobiology MS applications, the reagents enable quantitative profiling of glycan isoforms and discovery of glycan biomarkers; they provide improved ionization of labeled glycans for increased sensitivity and better retention of labeled glycans by reversed-phase liquid chromatography (RPLC)
• The aminoxyTMT reagents may be used to quantify a broad range of biologically important molecules including carbohydrates, steroids, or oxidized proteins
• Features: • Quantitative —enables relative quantitation of glycans or other carbonyl-containing biomolecules from multiple samples derived from cells, tissues or biological fluids • Stable —oxime bond and products formed by labeling reaction are stable and don't require an additional reduction step • Efficient —achieve labeling efficiency greater than 90% in one hour • Sensitive —20-fold increase in signal-to-noise compared to unlabeled native glycan MS analysis • Multiplex —able to identify and characterize up to six samples concurrently • Optimized —procedure and reagents optimized for excellent labeling efficiency and recovery of glycans Includes: • Label reagent sets contain sufficient aminoxyTMT tags for five sixplex experiments Requires: • PNGase F (or PNGase A) glycosidase and Waters Oasis™ HLB columns Applications: • Multiplex up to 6 different samples concurrently in a single experiment • Relative quantitation of glycans • Study of structural diversity of protein glycosylation • Study of glycosylation in cell signaling and regulation • Study of cancer progression, biomarker discovery and analysis of biotherapeutics Native glycans are difficult to study by mass spectrometry because of their poor ionization efficiency
• Quantitation of glycans is particularly challenging due to the lack of standards for all naturally occurring glycans and difficulties reproducibly quantifying multiple samples
• The aminoxy group has better reactivity with carbonyls and better stability of the labeled product compared to hydrazides
• Labeling with the aminoxyTMT reagents improves ionization of glycans, thus improving sensitivity, and enables relative quantitation of glycans for up to six samples concurrently
• These products are subject to a limited use label license.