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Proteomics Seminar Series 2012: Thermo Fisher Scientific
March 7, 2012 @ 11:30 AM - 1:00 PM
Utilization of enrichment methods and quantitative proteomics techniques to monitor cellular regulation
Speaker: Brian Reese, Ph.D., Proteomics Applications Specialist, Thermo Scientific, Pierce Protein Research
Mass spectrometry provides users with an unparalleled ability to look at cellular regulation in terms of monitoring both protein expression levels and post translational modification. However, current proteomics analysis is not without significant limitations caused by the dynamic range of proteins in a cell, the sensitivity and speed of instrumentation, and the need for contextual information for data analysis. By combining quantitative MS-analysis, using methods such as MRM/AQUA, SILAC, orTMT isobaric tags, with specific sample fractionation methods, the quality and depth for proteomic analysis can be greatly improved. Methods of sample fractionation described include enrichment of; phosphopeptides, kinases, sub-cellular organelles,cysteine containing peptides, and cell surface proteins. A novel approach for removing detergents from samples is also examined.
Exploring Quantitative Differences in Serum StimulatedHeLa CellLysates Using HybridOrbitrap Systems
Speaker: Craig P. Dufresne, Ph.D., Senior Technical Instructor for Mass Spectrometry, Thermo Fisher Scientific, Unity Lab Services
HeLacells cultured for 24 hours in low serum (0.5%) conditions and then serum stimulated (10%) prior to harvesting will be used to illustrate quantitative differences compared to normal growth conditions. Various methods for acquiring label freequantitation data on hybridOrbitrap high resolution mass spectrometers will be used. Discoveryquantitation using recursive base peak framing in SIEVE and targeted quantitation-high resolution extracted ion chromatograms in Pinpoint will be used for data processing and comparative presentations of the data.
Both seminars will be presented with lunch.
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