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
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