Dunn School | CPF (Headington) | CBRG | University of Oxford

Dunn School of Pathology

Central Proteomics Facility

Instruments - The Central Proteomics Facility has recently relocated to a custom built laboratory in the Sir William Dunn Pathology School.

For the last 5 years a Micromass Q-TOF Micro mass spectrometer has been the workhorse of the proteomics facility, however, this has now been replaced with a Thermo Orbitrap and an ABI 4800 MALDI TOF TOF mass spectrometer.

Orbitrap - A Thermo LTQ Orbitrap mass spectrometer coupled to a Dionex Ultimate 3000 nano HPLC system was installed in April 2007 and is now operational.

The Orbitrap is capable of mass resolving powers of up 100,000 (FWHM) and the mass accuracy is routinely better than 2 ppm. The instrument is capable of identifying proteins in the low fmol/high amol range (40% sequence coverage from 10 fmol BSA tryptic digest).

The Orbitrap is ideally suited to identifying multiple components in complex protein mixtures, identifying post translational modifications and comparative quantitation using SILAC.

4800 MALDI TOF TOF - An ABI 4800 MALDI TOF TOF mass spectrometer was installed in March 2007 and is now fully operational.

The 4800 is capable of mass resolving powers of 18,000 (FWHM) and the mass accuracy is routinely better than 50 ppm. The instrument is capable of identifying proteins in the low fmol/high amol. The instrument capable of LC MALDI and is equipped with a Dionex Ultimate 3000 nano HPLC system with Probot MALDI spotting system.

The 4800 is suited to rapidly identifying proteins in gel bands and 2D gel spots. By using LC MALDI the system is also capable of analysing complex mixtures and performing comparative quantitation using iITRAQ .

Liquid Chromatography and DIGE - In addition to mass spectrometry the Central Proteomics Facility is equipped with a range of liquid chromatographic systems, including an analytical flowrate HPLC and an AKTA purifier. The CPF also has DIGE (Differential In-Gel Expression) equipment, including gel tanks,Typhoon imager, DeCyder analysis software, spot picking robot etc (some equipment located in Physiology and some in Pathology). See the DIGE section of the website.

The CPF also has a Beckman PF2D 2-dimensional chromatography system for fractionating complex samples. The system separates intact proteins by their pI in the liquid phase in the first dimension, and by their hydrophobicity/hydrophilicity in the second dimension.

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