OPTIMIZATION OF PREPARATION OF MITOCHONDRIA FROM 25-100 MG SKELETAL-MUSCLE

A method for isolation of mitochondria from 25-100 mg skeletal muscle is described. The instrumental developments include a refined homogeni zation setup, special handling techniques, and equipment for biopsy st orage. The preparation medium was a standard ionic medium. All fractio ns were assayed for marker enzymes and the data used in optimization o f the yield. It was observed that the homogenization procedure excerts strong control on the integrity of the isolated mitochondria. The met hod was developed with pigeon breast muscle as the model tissue and us ed virtually unaltered for preparation from muscles of pigs, rats, and humans. The relative yield was 40-50% and the mitochondria were well coupled and showed high rates of phosphorylating respiration. (C) 1997 Academic Press.