https://www.selleckchem.com/products/iodoacetamide.html Mass spectrometry (MS)-based proteomics is currently the most successful approach to measure and compare peptides and proteins in a large variety of biological samples. Modern mass spectrometers, equipped with high-resolution analyzers, provide large amounts of data output. This is the case of shotgun/bottom-up proteomics, which consists in the enzymatic digestion of protein into peptides that are then measured by MS-instruments through a data dependent acquisition (DDA) mode. Dedicated bioinformatic tools and platforms have been developed to face the increasing size and complexity of raw MS data that need to be processed and interpreted for large-scale protein identification and quantification. This chapter illustrates the most popular bioinformatics solution for the analysis of shotgun MS-proteomics data. A general description will be provided on the data preprocessing options and the different search engines available, including practical suggestions on how to optimize the parameters for peptide search, based on hands-on experience.Two-Dimensional Polyacrylamide Gel Electrophoresis (2D-PAGE) provides two-dimensional maps where proteins appear separated according to their isoelectric point (pI) and molecular weight (MW). Usually these maps are very complex (i.e., hundreds or thousands of spots can be present in each map), and characterized by a low reproducibility, which hinders the possibility to identify reliable biomarkers unless robust methods are applied. The analysis of different sets of 2D-PAGE maps (e.g., control vs. pathological or control vs. drug-treated samples) to identify candidate biomarkers (proteins under- or over-expressed in different conditions) is usually carried out through image analysis systems providing a so-called spot volume dataset where each sample corresponds to a map described by the optical densities of all the detected spots. The identification of candidate biomarkers can be t