Proteomics Core Facility
We provide a full proteomics infrastructure for the identification and characterisation of proteins. This is centered around state-of-the-art mass spectrometry for MS and LC-MSMS experiments, and includes chromatographic and electrophoretic systems for protein and peptide separation.
Major projects and developments
- Molecular weight determination of intact proteins.
- Routine identification of proteins from coomassie and silver-stained gels.
- Identification of post-translational modifications.
- Nano-flow liquid chromatography coupled to high-resolution mass spectrometry: (LC-MSMS) for the identification of proteins in complex mixtures.
- Protein quantification by stable-isotope labelling (e.g. SILAC).
Services provided
Analysis of intact proteins:
- Molecular weight determination of intact proteins by ESI mass spectrometry (Bruker Maxis and Waters Q-tof).
- Determination of N- and C-termini of proteins and products of limited proteolysis.
- Protein separation by 1- and 2-dimensional electrophoresis using gel systems from 7 to 24 cm.
- Various gel staining protocols: Coomassie, silver, fluorescent staining by Sypro Ruby or Flamingo.
Proteomics:
- Pipeline for differential proteome analysis by 2D gels (incl. DIGE).
- Protein digestion in gel or in solution, using a variety of proteases.
- Enrichment of phosphopeptides (TiO2 and IMAC).
- Identification of post-translational modifi cations.
- Protein identification by MALDI peptide mass finger printing.
- Nano-flow reversed-phase chromatography (Dionex 3000 and Waters nano-Acquity systems) coupled in-line with ESI mass spectrometry.
- Ion trap (Bruker HCT) MS and MSMS for routine identification of proteins from coomassie and silver-stained gels.
- High-resolution and high mass-accuracy MS and MSMS (Thermo Orbitrap Velos) for identification and quantification of proteins in complex mixtures.
- Multi-dimensional peptide separation (isoelectric focusing and liquid chromatography);
- Protein quantification by stable-isotope labelling (SILAC and dimethyl labelling).
Technology partner
BIO-RAD