All proteomics services are performed on the Thermo Scientific Orbitrap Fusion tribrid mass spectrometer equipped with a Dionex UltiMate 3000 reverse-phase nano-UHPLC system.
For gel samples, excise the band of interest and cut it into approximately 1-2 mm2 pieces. Place these in an Eppendorf tube and submit. Please provide an approximation of protein amount and information on the stain used in the submission form.
For in-solution samples, we require a minimum of 10 ug of protein for analysis. PEG, glycerol and detergents are not compatible with mass spectrometric analysis and must be removed prior to sample submission. Please provide details on the buffer composition in the submission form.
As standard, we offer label-free quantitation (LFQ) as no specific sample pre-treatment is required. For LFQ, each sample is analyzed separately by LC-MS/MS and the results are subsequently compared across all samples to yield relative quantitation data.
Labelling of proteins for quantitation can be employed to enable multiplexed measurements (that is, samples are pooled together analyzed in a single LC-MS/MS experiment). There are several options for consideration:
Targeted quantitation of specific proteins for increased accuracy can be carried out by parallel reaction monitoring (PRM). PRM requires dedicated method development for each target and will thus be more time-consuming than other approaches.
Requirements are similar to those for protein identification; we need a minimum of 10 ug of protein for analysis. We recommend submitting three biological replicates for quantitation.
We offer the identification and localization of acetylation, methylation, phosphorylation, glycosylation and other modifications by LC-MS/MS. Analysis of post-translational modifications can be performed as part of qualitative or quantitative experiments. In certain cases, enrichment of modified peptides may be necessary prior to sample submission.
Other, non-routine experiments may be developed and offered on an individual basis. If you wish to start a new project, please contact Klaudia Kocurek to schedule a consultation prior to submitting samples.