title: Isolation and functional analysis of mitochondria from cultured cells and mouse tissue.
abstract: Comparison between two or more distinct groups, such as healthy vs. disease, is necessary to determine cellular status. Mitochondria are at the nexus of cell heath due to their role in both cell metabolism and energy production as well as control of apoptosis. Therefore, direct evaluation of isolated mitochondria and mitochondrial perturbation offers the ability to determine if organelle-specific (dys)function is occurring. The methods described in this protocol include isolation of intact, functional mitochondria from HEK cultured cells and mouse liver and spinal cord, but can be easily adapted for use with other cultured cells or animal tissues. Mitochondrial function assessed by TMRE and the use of common mitochondrial uncouplers and inhibitors in conjunction with a fluorescent plate reader allow this protocol not only to be versatile and accessible to most research laboratories, but also offers high throughput.
authors: Chemistry Department, Elon University.
authors: Thomas
authors: T
authors: Lampl
authors: Chemistry Department, Elon University.
authors: Jo A
authors: JA
authors: Crum
authors: Chemistry Department, Elon University.
authors: Taylor A
authors: TA
authors: Davis
authors: Department of Neurobiology and Anatomy, Wake Forest School of Medicine; Neuroscience Graduate Program, Wake Forest School of Medicine; ALS Center Translational Science Unit, Wake Forest School of Medicine.
authors: Carol
authors: C
authors: Milligan
authors: Chemistry Department, Elon University; vmoore3@elon.edu.
authors: Victoria
authors: V
authors: Del Gaizo Moore
mesh_terms: D000818:Animals; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008928:Mitochondria / Q000737:chemistry* / Q000502:physiology*