The Proteomics Shared Resource offers complete proteomic services and has considerable experience in identifying proteins from 1D and 2D gels using the current state-of-the-art electrophoresis and imaging equipment, robotic sample handlers and mass spectrometers for proteomic analysis. Core personnel also provide assistance to individual investigators with the design and implementation of proteomics studies.

The Proteomics Shared Resource provides complete proteomic support from start to finish. We offer cell lysis of cell pellets or protein extraction from cells, protein quantitation, protein preparation (precipitation). Following protein preparation from cells or tissue, protein separation services using 1D or 2D-PAGE techniques are available. The gels can be stained with various stains including Coommassie, Sypro Ruby, Deep Purple and Silver. Multiplex analysis by visualization of proteins and protein modifications using specialized fluorescent stains, and advanced gel image analysis are available to study phosphorylation, glycosylation and other modifications. Changes in protein expression levels are analyzed using Differential Image Gel Electrophoresis (DIGE) which uses cy dye labeling.

Protein spots of interest can be determined following excision from the polyacrylamide gel, protease digestion, and nano LC-MS/MS analysis of the peptides. If the protein identity cannot be ascertained by MALDI mapping, nano-LC/MS/MS is used to provide internal sequence of the protein. We have found this method to be sensitive and powerful, producing numerous sequences from 200 fmole of material and enabling us to identify proteins for which peptide mass mapping was unsuccessful. The Capillary LC system is capable of 2-D LC-chromatography and variable flow chromatography for the analysis and identification for complex protein mixtures (global proteomics.

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Mass Spectrometry and Proteomics Facility