Phospho-screen plus

Overview

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.

Steps Involved

• 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.

Deliverables

• 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.