Proteomics Facility
Dr Chris Storey will be retiring on 31st March
If you require full Biomolecular Analysis please contact David Knight david.knight@manchester.ac.uk 0161 2751561
Protein Identification Servicewithin the MIB
· Uses
- Identification of unknown proteins.
- Confirmation of the expression of cloned proteins.
- Detection of multiple proteins in precipitates or from binding experiments.
- Detection of proteins in biological fluids, for example CSF or serum.
This service is for the identification of proteins either from coomassie-blue stained gels, silver stained gels or from proteins in solution. The service is based on identification of tryptic peptides using nanoflow HPLC and ion-trap mass spectroscopy. The system is very sensitive - if you can see a band on a gel there is enough protein for an ID. As a bonus you also get some amino acid sequence from the identified peptides.
The service includes sample preparation by tryptic digestion, sample analysis by LCMS, data analysis and database searching.
The main instrument used to identify proteins is an LC packings nanoflow LC attached to a Bruker ion-trap mass spectrometer. The LC separates the tryptic peptides and sprays them directly into the mass spectrometer. This is set up to do automatic MS2/3 which will break the peptides in to smaller fragments and so give some amino-acid sequence data which is them used to identify the protein.
MALDI TOF Mass Spectrometry:
· Uses
- Accurate measurement of molecular mass.
- Looking at reaction products following chemical synthesis.
- Effects of heavy isotope labelling.
- Quality control.
We have an open access MALDI-TOF mass spectrometer. It can be used to measure the molecular weight of molecules from a few tens of Daltons up to 100,000 Daltons with minimal sample preparation. This mass spectrometer is a Kratos Axima-CRF and it can measure the mass of a 1000Da peptide to an accuracy of 0.02Da. Anyone in the MIB can use this machine after a couple of hours training.
If you are interested contact Eddie McKenzie edward.a.mckenzie@manchester.ac.uk or on 3064175
Other Mass Spectroscopic services in the MIB
There are a variety of other communal MS instruments available in the MIB for researchers to use:
Open Access ESI-TOF
GC-Trap
For information on these machines contact Reynard Spiess (phone 65157), Reynard.spiess@manchester.ac.uk, 1.034 or Paul Williams (phone 65185), Paul.Williams-2@manchester.ac.uk 1.034
For further information on using the protein identification service or using the open access MALDI please contact:
Eddie McKenzie edward.a.mckenzie@manchester.ac.uk or on 3064175