Acquisition of DIGE Instrumentation to Enhance Proteomic Research in Nevada

University Of Nevada Reno, Reno NV

Investigators

Abstract

DESCRIPTION (provided by applicant): This proposal is a resubmission of an NCRR Shared Instrumentation Grant originally submitted in March 2007. The Nevada Proteomics Center, in collaboration with INBRE and COBRE projects and NIH-funded investigators, requests equipment and software that will enable our 2-D Gel Laboratory to perform differential in-gel electrophoresis (DIGE) experiments. The requested items include: a Typhoon Trio laser imager, DeCyder v 6.5 and DeCyder Extended Data Analysis software, and a Genomics Solution ProPic II Spot Cutter with DIGE Upgrade. Two-dimensional (2-D) gels are currently the principle method for separating and quantifying proteins in proteomics experiments. However, standard 2-D gels are notoriously inconsistent and gel-to-gel variations make accurate determinations of change in protein abundance difficult. DIGE technology overcomes these obstacles by running a control and experimental sample, along with an additional normalizing sample, on the same 2-D gel. Each of these 3 types of samples is labeled with a different fluorescent dye;these dyes are detected by the Typhoon imager and differences in dye intensities are quantified by DeCyder software. Protein spots of interest will be cut from the gel by the ProPic II spot cutter and identified by mass spectrometry. The requested system represents the state-of-the-art in DIGE technology and integrates well with our existing instrumentation. Our 2-D Gel Lab, which was set up with BRIN and EPSCoR funding, has 5 years of experience plus all necessary equipment for generating the gels. The DIGE technology would be invaluable for the research of many of our NIH-funded investigators at University of Nevada who are attempting to detect physiologically important protein changes in a variety of disease states. PUBLIC HEALTH RELEVANCE: While all cells in any one organism contain the same DNA, the proteins that are made from this DNA vary greatly from one cell to another and vary in response to environmental influences or disease states. By studying changes in the amounts or chemical modifications of proteins in a cell, we can better understand the molecular basis of a disease and how to treat it. In this grant proposal, we request funds to purchase a sensitive and accurate way to compare the amounts of proteins produced in a cell under healthy and disease states.