Ubiquitination is an essential regulatory modification that controls protein turnover. Our workflow identifies ubiquitination sites through the diGly remnant left on modified lysines after trypsin digestion, and quantifies them proteome-wide across multiplexed samples.
At a glance
- Thousands of ubiquitination sites quantified per study
- High-affinity enrichment with PTMScan technology
- TMT 18-plex with RTS-MS3 on Orbitrap Eclipse instruments
- Suited to protein turnover studies and PROTAC specificity screening, in cell lines and tissues
How it works
Protein extracts from up to 18 samples are digested with trypsin, and ubiquitinated peptides are immunoenriched through their characteristic diGly remnant. The enriched peptides are labeled with TMT, mixed and analyzed by RTS-MS3 on Orbitrap Eclipse instruments, for sensitive, reliable measurements across thousands of sites with no missing values.
Example data: ten mouse tissues
Ten mouse tissue samples were profiled together in a single 10-plex experiment, quantifying 4,730 ubiquitination sites across 1,984 proteins. Each tissue type shows a distinct ubiquitination signature, reproduced closely across biological replicates.
Spotlight: individual sites
Three sites show what the data captures. Neuromodulin sites agree closely between replicate brain samples. Ubiquitination of myosin light chain 3 is specific to heart, measured across a wide dynamic range. Subtle differences in 60S ribosomal protein L4 are quantified reproducibly.
