University of Bristol Proteomics Facility |
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Protein Identification
From complex mixtures
Proteins are separated using 1D gels and the gel lane cut into a number of slices. Each slice undergoes in-gel tryptic digestion using our ProGest automated digestion unit which minimises sample handling and hence keratin contamination. The resulting peptides are then analysed by RP nano-LC MSMS using our LTQ-Orbitrap Velos mass spectrometer.
Alternatively, protein samples in solution can be digested with trypsin and then fractionated by Strong Cation eXchange (SCX) chromatography. Each SCX fraction is then analysed by RP nano-LC MSMS as above.
For each approach, the mass spectral data from each fraction is combined prior to database searching using an in-house Mascot server to identify the proteins present in the sample. The final Mascot report is emailed to the client.