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 analyses 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 180 polar metabolites spanning 32 different classes, including metabolites involved in central carbon metabolism, tryptophan catabolism, and amino acid metabolism; 2) metabolic flux analysis using stable isotope labeled tracers; 3) untargeted polar metabolite quantitative comparisons, and 4) untargeted lipidome quantitative comparisons. Services 3 and 4 are being validated and have limited availability on a trial basis.
- 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 up to approximately 180 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
- Flux (isotope tracers) analysis of cell metabolism using LC-MS
- Untargeted polar metabolite quantitative comparisons
- Untargeted lipidome quantitative comparisons
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 Wistar Institute Cancer Center.
The Wistar Institute
David W. Speicher, Ph.D.
Hsin-Yao Tang, Ph.D.
Wistar Research Assistants