This service enables a global view of protein and site specific phosphorylation events using a workflow optimized for a 24hr (8 x 3hr) LC-MS/MS analysis. This workflow can be used to study phosphorylated species from complex samples such as cell lysates, tissues and biofluids. Samples are processed by trypsin digestion, peptides are fractionated using a basic reverse phase strategy. Phosphorylated peptides are enriched using titanium dioxide microparticles. This service uses 24hours of instrument time versus the 2hrs used for Phospho-screen. The increased instrument time is reflected in the number of phospho protein/peptide identifications.
- Sample preparation. Protein extraction, solution digestion with trypsin, basic reverse phase fractionation, pooling of fractions using a custom matrix and enrichment of phosphopeptides with titanium dioxide.
- LC-MS/MS. 8x3hr LC-MS/MS using a Q Exactive HF or Fusion Lumos.
- Data analysis. Phosphorylation site identification quantitation with MaxQuant. Differential analysis with Perseus.
- Report generation: PDF/Excel
The turnaround time is 2-3 weeks.
- Written Report containing details of your samples, our methods and a summary of the data.
- An Excel workbook containing the detailed qualitative and quantitative protein identification data.
- A Scaffold file to enable your own deeper data mining.
An example report is available upon request.