期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 97, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/52076
关键词
Cellular Biology; Issue 97; Mitochondria; TMRE; cytokines; ALS; HEK cells; fluorescence; mitochondrial dysfunction; mitochondrial membrane potential; cytochrome c
资金
- NSF [CHE-1229562]
- Office of Undergraduate Research at Elon University
- Elon Chemistry Department
- Elon Lumen Prize
- Elon College Fellows Program
- Elon College Honors Program
- Division Of Chemistry
- Direct For Mathematical & Physical Scien [1229562] Funding Source: National Science Foundation
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.
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