Proteomics & Metabolomics Facility
The Proteomics and Metabolomics Shared Resource provides high sensitivity proteomics and metabolomics analyses using state-of-the-art mass spectrometry instruments and methods. Consultation with facility staff concerning experimental design and sample preparation is recommended prior to sample preparation to ensure optimal experimental design.
Proteomics services include: 1) quantitative, in-depth global comparisons of sub-proteomes, complete proteomes, and secretomes using integrated ion current, SILAC or TMT labeling; 2) global quantitative comparisons of posttranslational modifications (PTMs) such as ubiquitination, acetylation, or phosphorylation; 3) detailed characterization of individual purified proteins including PTMs; 4) identification of components in protein complexes (e.g. pull-downs) including estimation of stoichiometries (where appropriate); 5) characterization of intact protein and peptide masses using either MALDI-MS or ESI-MS; and 6) HPLC peptide mapping with UV detection.
Metabolomics services include analysis of polar metabolites or lipids extracted from cells, biological fluids, conditioned media, or tissues. Specific services include: 1) targeted relative quantitation of approximately 200 polar metabolites spanning 32 different classes; 2) 13C stable isotope tracer analysis; 3) untargeted polar metabolite quantitative comparisons, and 4) untargeted lipidomics for quantitative profiling of global lipids or acylceramides (after mild saponification), and 5) targeted relative quantitation of free fatty acids, total fatty acids (after saponification), and eicosanoids, including prostaglandins and HETEs. Samples for all these applications are analyzed using the Thermo Q Exactive HF-X mass spectrometer. Metabolites are separated using HILIC chromatography, global lipids and acylceramides are separated on a C30 reversed-phase column, and fatty acids and eicosanoids are separated on a C18 column.
- In-gel protease digestion using Colloidal Coomassie or MS-compatible silver stained gels
- In-solution protease digestion
- LC-MS/MS for protein identification
- High resolution accurate mass ESI-MS analysis of small intact proteins
- MALDI-MS protein and peptide analysis
- 1D SDS-PAGE sample preparation and fractionation
- Reverse phase microbore HPLC peptide mapping
- Targeted relative quantitation of approximately 200 validated metabolites
- Characterize PTMs (purified proteins or global profiling)
- Global label-free quantitative analysis of proteomes using Gel/LC-MS/MS
- Quantitative comparisons using SILAC or isobaric tags (TMT)
- High pH protein fractionation for LC/LC-MS/MS
- Analysis of protein-protein crosslinks
- Targeted quantitation of selected proteins using PRM-MS of prototypic peptides
- 13C stable isotope tracer analysis of cell metabolism using LC-MS.
- Untargeted polar metabolite quantitative comparisons
- Untargeted lipidomics for quantitative profiling of global lipids and acylceramides
- Targeted quantitation of free fatty acids, total fatty acids, and eicosanoids
Click here for gel shipping instructions and SDS-gel guidelines.
For pricing information, visit iLab or contact the managing director.
This facility is supported in part by a Cancer Center Support Grant (CCSG) awarded by the National Cancer Institute (NCI) to the Ellen and Ronald Caplan Cancer Center.
Click here for information on how to acknowledge our Shared Resources in your publications and grants.
The Wistar Institute
Hsin-Yao Tang, Ph.D.
Aaron R. Goldman, Ph.D.
Wistar Research Assistants