Proteomics Core FacilityServices
The Proteomics Core Facility offers a range of techniques for the identification and characterization of proteins. These include the following, often used in combination – please enquire for additional applications that may fit your needs.
Protein and Peptide Separation
- Iso-electric focusing of proteins and peptides, using a wide range of pH gradients. Proteins can be separated as a first step in 2D gel electrophoresis. Peptide IEF (using the OFFgel fractionators) is a relatively new addition which is typically used as a pre-fractionation step in peptide-based proteomics prior to LC-MSMS.
- SDS-PAGE gel electrophoresis on gels ranging from 7 to 24 cm. Either in 1D or 2D gel electrophoresis.
- Various staining protocols: Coomassie, silver, fluorescent staining by Sypro Ruby or Flamingo, ProQ-Diamond phosphostain.
- Densitometry of coomassie and silver stained gels.
- Laser fluorescence imaging of fluorescently stained gels.
- PDQuest software for differential analysis of 2D gels.
- Differential analysis by DIGE, including fluorescent staining, quantitation and statistical analysis of 2D spot patterns.
- Automatic excision of spots from 2D gels
Protein digestion in gel or in solution, using a variety of proteases (trypsin, LysC, ArgC, chymotrypsin)
Peptide purification using ZIP tips or Stage-tips.
Mass Spectrometry: MS, MSMS and LC-MSMS
- Protein identification by MALDI peptide mass fingerprinting
- ESI-MS for protein molecular weight determination under denaturing or non-denaturing conditions.
- Nano-flow reversed-phase chromatography coupled in-line with ESI mass spectrometry (ion trap / MaXis Q-Tof / LTQ Velos).
- Ion trap MS and MSMS for routine identification of proteins from coomassie and silver-stained gels.
- Peptide fragmentation by CID and ETD.
- High-resolution and high mass-accuracy MS and MSMS using MaXis Q-Tof / LTQ Velos for detailed characterisation of proteins, including post-translational modifications.
- Identification of phosphorylation and other post-translational modifications.
- Protein identification from peptide fingerprints and LC-MSMS using Mascot. Databases can be customized to the user’s needs.
- Reporting to users in appropriate detail - from protein-summary in Excel to spectra with peak annotation.
Price List (EMBL internally only)