Proteomics and Metabolomics Facility

Overview

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. 

Services

Routine:

  • 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

Specialized:

  • 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 MRM-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.

Pricing information can be found on 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.

Contact Us

The Wistar Institute
Proteomics and Metabolomics Facility, Room 252
3601 Spruce Street
Philadelphia, PA 19104

Phone: 215-898-3830/3181
Fax: 215-898-0664

Facility Hours:
9:00 a.m.-5:00 p.m., Monday-Friday

David W. Speicher, Ph.D.
Scientific Director

Hsin-Yao Tang, Ph.D.
Managing Director

Thomas Beer
Nicole Gorman
Wistar Research Assistants