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PROTEOMICS

PROTEOMICS

Introduction to High Performance Liquid Chromatography

Gel-Free/FASP Digestion

High Performance Liquid Chromatography (HPLC) is an essential part of Pro- teomics workflows. It is usually used to fractionate complex protein/peptide mixtures, in order to increase proteomic coverage or improve detection of post-translational modifications such as phosphorylation. In this course, participants will be given an introduction to HPLC and learn how to start the HPLC system. They will become familiar with the equipment: pump unit and fraction collector. Upon completion, users will be able to prepare HPLC buffers for peptide fractionation; set up the control program to run a peptide frac- tionation experiment; and perform post processing of samples and storage. PARTICIPANT PROFILE This course is intended for students and research professionals from KAUST, and their collaborators from academia and industry, who want to perform shot- gun proteomics using off-line HPLC fractionation. PREREQUISITES • Active Badger account

This training introduces Filter Aided Sample Prep (FASP) as a method for gen- erating tryptic peptides from crude lysates for subsequent LC-MS analysis. This method allows the analysis of protein mixtures generated from detergent-lysed cells and tissues. It is thus suitable for studying whole proteomes and membrane rich fractions. As the name indicates, the essential steps of the protein clean-up and digestion are carried out using a filtration device. This course will cover the key features of FASP and subsequent peptide preparation for LC-MS analysis. Participants will learn how to process lysates prepared using detergents, be- come familiar with protein digestion using filtration devices, and learn peptide cleanup for MS analysis. PARTICIPANT PROFILE This course is intended for students and research professionals from KAUST, and their collaborators from academia and industry, who want to perform FASP digestion for the analysis of detergent-based lysates. PREREQUISITES • Active Badger account

DURATION 3 hours FREQUENCY Twice monthly

DURATION 1 day FREQUENCY Twice monthly

• Laboratory Safety Training • Hazardous Waste Training • Emergency Incident Preparedness Training MAXIMUM PARTICIPANTS 3

• Laboratory Safety Training • Hazardous Waste Training • Emergency Incident Preparedness Training MAXIMUM PARTICIPANTS 3

Learn Develop Excel

Specialized Instrument & Method Training Catalog

Edition 1, Sep te mber 2020

117

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