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Course - Cold Spring Harbor, NY, United States

 

This intensive laboratory and lecture course will focus on cutting-edge proteomic approaches and technologies. Students will gain practical experience purifying and identifying protein complexes and posttranslational modifications. In a section focused on quantitative whole proteome analyses or top-down proteomics, students will gain hands-on experience using two-dimensional gel electrophoresis and mass spectrometry analysis. Students will use differential in-gel electrophoresis (DIGE) for gel-based protein quantification. Differentially expressed proteins will be recognized by statistical methods using advanced gel analysis software and identified using MALDI mass spectrometry. For shotgun proteomic analysis sections or bottom-up proteomics, students will use label-free and covalent isotopic-labeling quantitative approaches to differentially profile changes in protein complexes and whole proteomes. Students will be trained in high-sensitivity microcapillary liquid chromatography coupled with nanospray-ESI and tandem mass spectrometry analysis. Students will learn both single-dimension and multidimensional separation methods. In a section focused on targeted proteomics, students will learn to analyze and process shotgun proteomic data for the development of SRM/MRM assays that accurately identify and quantify targeted proteins. Students will design transitions for selected peptides and perform SRM/MRM mass spectrometry assays. They will learn to process and interpret the acquired data to measure changing quantities of targeted proteins in a variety of biological samples. For all sections of the course, a strong emphasis will be placed on data analysis. A series of outside lecturers will discuss various proteomics topics including: imaging by mass spectrometry, de novo sequence analysis, advanced mass spectrometry methods, protein arrays, and functional proteomics. The aim of the course is to provide each student with the fundamental knowledge and hands-on experience necessary for performing and analyzing proteomic experiments. The overall goal is to train students to identify new opportunities and applications for proteomic approaches in their biological research.