Equipment
AB Sciex QSTAR Elite Quadrupole-TOF mass spectrometer with Nanospray source and Tempo nanoLC system
Through the utilisation of nanoflow chromatography (~0.3 microlitre/minute flow rates), extremely small chromatography columns (75 micrometre internal diameter) and nanoelectrospray ionisation, this LC/MS/MS system can identify proteins from sub-nanogram amounts of protein. We routinely analyse and identify peptides from 2D PAGE fractionated protein spots (low complexity samples) through to in-solution proteolytic digests of intact cells (high complexity samples).
AB Sciex 5800 MALDI TOF/TOF mass spectrometer
Using the alternative ionisation method of Matrix Assisted Laser Desorption/Ionisation, this instrument is used to identify proteins from 2D PAGE fractionated protein spots and to perform Imaging Mass Spectrometry analysis on tissue sections and bacterial colonies.
Agilent 1200 capillary HPLC system
For peptide samples that are too complex for LC/MS/MS, this system is used to fractionate peptide and protein samples by orthogonal chemical properties. Fractionated samples are then subjected to LC/MS/MS on the QSTAR systems.
Bio-Rad Protean IEF Cells, Criterion Gel System and Protean XL Gel System and Proteome Systems IsoelectrIQ isoelectric focusing system
2D PAGE is still the highest resolution technique for the separation of proteins and their isoforms, while 1D SDS PAGE is a robust fractionation technique prior to LC/MS/MS for complex proteome samples. Our electrophoresis equipment can perform size fractionation of 26 samples on a single gel in ~30 minutes or 2D PAGE in 7cm, 11cm or 18cm formats using pre-cast or self cast gradient gels.
Bio-Rad 491 PrepCell, MiniPrepCell, RotorFor and MicroRotorFor preparative electrophoresis systems
Utilised for prefractionation of complex protein samples by either size or charge to reduce complexity. When combined with narrow pH range immobilised pH gradient strips, the depth of proteome coverage using 2D PAGE is dramatically increased.
Bio-Rad PharosFX Plus Fluorescent and ChemiDoc XRS Gel Imaging Systems, and PDQuest and Proteomeweaver 2-D Gel Analysis Software
Gel based proteomics requires imaging systems to visualise band and spot patterns, and software to determine what proteins are being differentially expressed. Our imaging platforms are able to image a range of fluorescent and visible protein gel stains while our gel analysis software allows us to determine which spots are changing so that they may be excised from the gel and the protein identified by mass spectrometry.
Proteome Systems XCise gel spot excision robot and digestion platform
For the imaging of coomassie blue stained gels, automated cutting of gel spots, automated proteolytic digestion of proteins within gel spots, automated desalting of digests and spotting of peptides onto MALDI targets for analysis by mass spectrometry.
