Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC)-Based Quantitative Proteomics and Phosphoproteomics in Fission Yeast
| Autor: | Weronika E. Borek, Kenneth E. Sawin, Boris Macek, Claudia C. Bicho, Alejandro Carpy, André Koch, Silke Hauf |
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| Rok vydání: | 2017 |
| Předmět: |
Proteomics
0301 basic medicine Proteome 030102 biochemistry & molecular biology Phosphopeptide Chemistry Quantitative proteomics Phosphoproteomics Phosphoproteins Mass spectrometry Orbitrap Mass Spectrometry General Biochemistry Genetics and Molecular Biology Yeast law.invention Fungal Proteins 03 medical and health sciences 030104 developmental biology Biochemistry law Isotope Labeling Stable isotope labeling by amino acids in cell culture Schizosaccharomyces Amino Acids |
| Zdroj: | Cold Spring Harbor Protocols. 2017:pdb.prot091686 |
| ISSN: | 1559-6095 1940-3402 |
| DOI: | 10.1101/pdb.prot091686 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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