on 07-08-201309:26 AM - edited on 10-15-202110:59 AM by Closed Account
Viner R, Bomgarden R, Blank M, Rogers J. ASMS 2013 Poster Note Purpose: To develop Tandem Mass Tag™ (TMT™) 10-plex reagents by combining
current Thermo Scientific™ TMTsixplex™ reagents with four TMT6 reagent isotope
variants on multiple high-resolution mass spectrometry platforms.
Methods: HeLa cell lysates and bovine serum albumin (BSA) digests were labeled
with Thermo Scientific™ TMT10plex™ reagents. Aliquots from six, eight or all ten
channels were mixed in different ratios and analyzed on three different Thermo
Scientific™ Orbitrap™-based instruments.
Results: We have extended the multiplexing capabilities of TMT reagents from 6 to 10
without increasing the size or structure of the tag by utilizing the 6 mDa mass
difference between 13C and 15N isotopes of the mass tag reporter ions. Mass
spectrometry analysis on three different Orbitrap-based instruments using FT
MS2/MS3 shows no negative effect on the number of protein identifications or quality
of quantification in a complex digest by increasing sample multiplexing from 6 to 10.