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In addition to the physiological functions of protein-bound lipoic acid, there is increasing scientific and medical interest in potential therapeutic uses of pharmacological doses of free (unbound) lipoic acid (3)

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Protein Extraction and Immunoblot Analysis To analyze the levels of specific mitochondrial proteins in the muscle, 2 mg of lyophilised muscle was homogenized in ice-cold buffer (100 lmg 1 dry weight) consisting of 2 mM HEPES (pH 7.4), 1 mM EDTA, 5 mM EGTA, 10 mM MgCl 2 , 50 mM -glycerophosphate, 1% TritonX-100, 2 mM dithiothreitol, 1% phosphatase inhibitor cocktail (Sigma P-2850) and 1% (v/v) Halt Protease Inhibitor Cocktail (Thermo Scientific, Rockford, United States) using a Bullet Blender (NextAdvance, New York, United States)