Stable Isotope Labelling with Amino Acids in Cell Culture – Size Exclusion Chromatography – Mass Spectrometry

Most proteins execute their functions through interacting with other macromolecules, and protein complexes formed by organized protein-protein interactions represent a primary functional module in the cells. Recently, People combine several proteomic techniques including cell fractionation, protein chromatography, and quantitative mass spectrometry (MS) to analyze the study composition and dynamics of protein complexes. To quantify the change of protein complex by this method, we combine it with the SILAC technique to detect the difference between different isotope-labeling samples. Here we harvest Saccharomyces cerevisiae cells in heavy, medium, and light labeling mediums. Then, extract the protein from cell pellets and separate it into 27 fractions by Size-exclusion chromatography. Using LC-MS/MS to detect protein from each fraction for doing protein complexes analysis