The Mass Spectrometry Shared Resource provides services to elucidate the primary structure of novel proteins and to analyze protein/protein interactions, post-translational modifications and protein levels. The Resource also offers the ability to detect lipids, metabolites, and other small molecules. The equipment in the facility has been upgraded to provide state-of-the-art analysis using microcapillary LC-MS/MS on linear ion traps and Orbitraps, hybrid quadrupole-TOF mass spectrometers and MALDI-TOF.
|Shared Resources Staff
|Darryl Pappin, Ph.D.
Facility staff is available to discuss project design and progress. Contact Keith Rivera to schedule an appointment. Services include:
- Quantitative global proteomic profiling using iTRAQ and SILAC, including data analysis, merging of separate datasets and statistical treatment of results.
- Quantitative cysteine profiling for global redox proteomics.
- GSH/GSSG measurement (MRM) for redox assessment.
- Multiple reaction monitoring (MRM) assays for:
- Phosphatidylinositols (PIs) including head group, chain length, degree of saturation and phosphorylation state for 20 different PI lipid species.
- Small molecule/drug measurement & clearance in serum & tissues (e.g. Clozapine and Clozapine N-Oxide, Vitamin C, Vitamin K).
- Lipid Analysis, including sample preparation and extractions, MRM assay development, LC-MS/MS and data analysis.
- Quantitative iTRAQ for protein complex IP.
- Quantitative metabolite profiling of ~250 core metabolites.
- Intact protein mass measurement for drug binding and to support crystallographic studies.
- Training in LCMS and MS/MS, both for instrument operation and data analysis.
- Consultation in experimental planning and customized experimental design.
The Shared Resource currently has four mass spectrometers in operation:
- Thermo Scientific Linear ion trap-orbitrap (Velos Pro): iTRAQ screens, protein identification, protein complexes (iTRAQ), post-translational modification analysis (PTM, including phospho, acetyl, ubiquitination, glycosylation and oxidation).
- Thermo Scientific Linear ion trap-orbitrap (XL): protein ID, PTM analysis and intact mass measurement.
- Thermo Scientific Vantage Triple Quadrupole mass spectrometer (TSQ): peptide MRM, lipids, carbohydrates, small molecules & metabolites
- Thermo Finnigan Linear ion trap (LTQ).
Sample preparation for the instruments is supported by a number of capillary liquid chromatography systems from Agilent and Proxeon.
Proteomics data is analyzed with the following data analysis packages:
- Mascot and Mascot Integra