300402 UE Mass Spectrometry based Proteomics (2015S)

We offer a practical university course, which provides training in mass spectrometry (MS)-based proteomics. In this laboratory course (limited to 12 students) we will introduce general concepts of MS based proteomics and discuss criteria that are required to prepare protein samples of sufficient quality and purity for MS analysis. You will perform sample preparation techniques (including affinity purification of proteins, reduction and alkylation of cysteines, digestion with trypsin), followed by RP-HPLC (reversed phase high-pressure liquid chromatography) MS measurement - the operation of the mass spectrometer will be demonstrated. You will also be introduced to MS data analysis using freeware MS data analysis tools such as MaxQuant, OpenMS and Perseus. We will furthermore aim to quantify changes in the post-translational modification pattern of the analyzed protein by applying label free quantification methods.