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  • Title: Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC)-Based Quantitative Proteomics and Phosphoproteomics in Fission Yeast.
    Author: Carpy A, Koch A, Bicho CC, Borek WE, Hauf S, Sawin KE, Maček B.
    Journal: Cold Spring Harb Protoc; 2017 Jun 01; 2017(6):pdb.prot091686. PubMed ID: 28572185.
    Abstract:
    Modern mass spectrometry (MS)-based approaches are capable of identifying and quantifying thousands of proteins and phosphorylation events in a single biological experiment. Here we present a (phospho)proteomic workflow based on in-solution proteome digestion of samples labeled by stable isotope labeling by amino acids in cell culture (SILAC) and phosphopeptide enrichment using strong cation exchange (SCX) and TiO2 chromatographies. These procedures are followed by high-accuracy MS measurement on an Orbitrap mass spectrometer and subsequent bioinformatic processing using MaxQuant software.
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