title: Stable isotope labeling for proteomic analysis of tissues in mouse.
abstract: Since the first metabolic labeling experiments with stable isotopes beginning of the last century, several approaches were pursued to monitor protein dynamics in living animals. Today, almost all model organisms from bacteria to rodents can be fully labeled with SILAC (stable isotope labeling of amino acids in cell culture) amino acids. The development of special media and diets containing the labeled amino acids provides an efficient way to metabolically label prokaryotic and eukaryotic organisms. Preferentially, the essential amino acid lysine ((13)C6-lysine) is used to label mice (Mus musculus) and after one generation the natural isotope is fully replaced by the stable (13)C6-lysine isotope. So far, the SILAC mouse approach has been used to analyze several transgenic and knockout mouse models. Spike-in of labeled proteins into non-labeled samples provides an accurate relative protein quantification method without any chemical modification. Here we describe how to establish a SILAC mouse colony and describe the analysis of skeletal muscle tissue with different metabolic and contractile profiles.
authors: Max Planck Institute for Heart and Lung Research, Lugwigstr. 43, 61231, Bad Nauheim, Germany.
authors: Soraya
authors: S
authors: Hölper
authors: Aaron
authors: A
authors: Ruhs
authors: Marcus
authors: M
authors: Krüger
mesh_terms: D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry; D005260:Female; D007553:Isotope Labeling / Q000379:methods*; D008239:Lysine / Q000737:chemistry*; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D009124:Muscle Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism*; D059748:Proteolysis; D040901:Proteomics / Q000379:methods*
chemical_list: D002247:Carbon Isotopes; D009124:Muscle Proteins; D008239:Lysine