PROTEOMICS FACILITY

Duke University, Durham NC

Investigators

Linked publications, trials & patents

Abstract

The main objective of the Proteomics Facility is to provide state of the art technology for the identification and characterization of proteins for Cancer Center members. A second goal is to carry out this work in a timely manner and at an affordable price that would not be prohibitive to complex protein mixture characterization. A third goal is to educate the investigator on how to get the best use out of the technology in the core. This includes discussion on techniques in protein separation, enrichment and staining techniques. Based on these goals the facility was equipped with an Applied Biosystems 4700 MALDI TOF TOF mass spectrometer (._M_atrix Assisted Laser Desorption Ionization Time of Flight Time of Flight), an ABI QSTAR TOF mass spectrometer and two ABI Edman automated sequenators. Other equipment include a Genomic Solutions robotic tryptic digestion and MALDI target spotting station, a Waters microbore HPLC and Amersham Pharmacia SMART system, a Macintosh X server work station for in house data base search with FASTS algorithm, a offline Dell workstation for MASCOT or PEPSEQUEST searches. An important aspect of the core expertise and capabilities are based upon the perceived needs of the individual investigators at Duke. The majority of users of the core hold peer reviewed grants centered on hypothesis driven research projects. Investigators of this type tend to have specific questions in mind and typically the type of work given to the facility involves identification of one or more proteins in an SDSPAGE gel. Typically gels are derived from experiments involving an antibody or recombinant protein pull down from a cell extract, followed by protein staining, isotope or fluor tagging. The investigator will indicate which proteins are of interest for protein identification. The numbers of proteins for identification for any one given job can vary from 1 to 100. Other projects (<10%) generally involve protein characterization, such as phosphorylation or glycosylation site analysis or recombinant protein analysis. The facility has limited capacity at the current time to handle high throughput proteomic analysis projects, such as proteomic analysis of multiple serum or tissue samples derived from patients. Although, there have been no requests for this capability, we anticipate future interest and will seek to expand the cores capabilities to meet this foreseen demand.