Global Protein Quantification of Mouse Heart Tissue Based on the SILAC Mouse

摘要: Metabolic labeling of living organisms with stable isotopes has become a powerful tool for global protein quantitation. The SILAC (stable isotope amino acids in cell culture) approach is based on the incorporation nonradioactive-labeled isotopic forms into cellular proteins. effective immortalized cells and single-cell (e.g., yeast bacteria) was recently extended to more complex organisms, including worms, flies, even rodents. administration (13)C6-lysine (heavy) containing diet one mouse generation leads complete exchange natural (light) (12)C6-lysine. SILAC-labeled are mainly used as heavy "spike-in" standard nonlabeled counterparts, combination high-performance mass spectrometers allows proteomic screening. Here we fully labeled mice identify proteins pairs from isolated cardiomyocytes, analyzed β-parvin-deficient hearts. Our confirmed absence β-parvin revealed simultaneously clear up regulation α-parvin heart tissue. In this protocol, describe colony show two approaches perform proteome-wide analysis Thus, spike-in readily available procedure straightforward systematic disease models knockout mice.