Gel electrophoresis-based proteomics of senescent tissues.
Cellular aging is a fundamental biological process, and mass spectrometry-based proteomics has been widely used for the global identification of age-related changes in a variety of tissues. The proteomic profiling of senescent skeletal muscles has revealed a variety of alterations in proteins associated with the contractile apparatus, cell signaling, ion homeostasis, metabolism, and the cellular stress response. Here, we outline the two-dimensional gel electrophoretic separation and fluorescent labeling of the urea-soluble protein complement from aged diaphragm muscle. This chapter describes the various experimental steps involved in gel electrophoresis-based proteomics, including protein extraction, isoelectric focusing, slab gel electrophoresis, fluorescence labeling, image analysis, protein digestion, mass spectrometric identification of proteins and immunoblotting.
Department of Biology, National University of Ireland Maynooth, Maynooth, Kildare, Ireland.
Steven
S
Carberry
Kay
K
Ohlendieck
D000375:Aging / Q000502:physiology; D000818:Animals; D016922:Cellular Senescence / Q000502:physiology; D003964:Diaphragm / Q000166:cytology / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D051379:Mice; D009124:Muscle Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012428:Ruthenium / Q000737:chemistry; D055948:Sarcopenia / Q000378:metabolism*
D009124:Muscle Proteins; D012428:Ruthenium