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Increasing the Multiplexing of Protein Quantitation from 6- to 10-Plex with Reporter Ion Isotopologues

Reputable Mentor II
Reputable Mentor II
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.

Thermo Fisher Scientific
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Last update:
‎10-15-2021 10:59 AM
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