Better Ion Transmission With Segmented Quadrupole

Segmented Quadrupole

•Improved transmission across m/z range and for narrow windows

•Brighter Source and Segmented Quad allows the use of a 0.4 amu isolation without loss of IDs (here for 1 ug HeLa, DD OT IT CID, 2 h runs, n=2)

•Improved performance for TMT quantitation

•Improved performance for PRM and DIA

•Improved performance for top down

Improved TMT SPS MS³ Performance

TMT Dilution

•Standard HeLa digest, labeled with TMT0 analyzed with an 85 min gradient using SPS-MS3

•Sample diluted 1:1, 1:10, 1:30, 1:50, 1:100

•The number of MS3 acquisitions was similar in both analyses

•The number of unique peptides quantified was systematically higher with the Orbitrap Fusion Lumos MS

Improved Low Level Quan: Ubiquitinated Peptides

TMT10 Quantitation of Ubiquitinated Peptides

•Human HTC116 cells were treated with a proteasome inhibitor (Bortezomib) for 16 h and analyzed with TMT 10-plex (5 treated vs. 5 untreated)

•Two fractions were prepared •With higher amount

•With lower amount

•25-73% more quantifiable peptides

ASMS Lecture: Rose et al. Isobaric labeling enables 10-Plex quantitative analysis of ubiquitylated peptides: A diagnostic ion to improve identification and quantification

SPS MS³ Quantification on Orbitrap Fusion Lumos MS

Results: Best possible accuracy and precision by reducing co-isolated interferences.

TMT Technology is More Precise than Label Free Quan

“We compared the average and median CVs (calculated for the whole dataset containing ca. 4000 proteins quantified with ≥2 peptides) between the three biological replicates of the same treatment. Ignoring the fact that the cell lines were different, the results are clearly in favor of TMT. In other words, TMT produced two times lower CVs than our label-free quantification, which we thought was pretty good. I am stunned…”

TMT Used for Protein Research in…

Straightforward Workflow

Result: Complete software and method development suite from reagents to data analysis