期刊
MECHANISMS OF AGEING AND DEVELOPMENT
卷 124, 期 1, 页码 33-41出版社
ELSEVIER IRELAND LTD
DOI: 10.1016/S0047-6374(02)00167-7
关键词
2-D PAGE; mitochondria proteins; aging; skeletal muscle
资金
- NATIONAL INSTITUTE ON AGING [P30AG013319, P01AG020591, P01AG014674, P01AG019316] Funding Source: NIH RePORTER
- NIA NIH HHS [P01 AG20591, P01AG14674, P01 AG19316, P01 AG13319] Funding Source: Medline
To study the effect of aging and anti-aging strategies on mitochondria, we have characterized a two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) system to analyze the profile of mitochondrial proteins. We have optimized the separation of proteins by 2-D PAGE and established the linearity and reproducibility of the system with mitochondria isolated from skeletal muscle of mice. Using total mitochondria protein ranging from 10 to 200 mug, we found that 74% of the proteins resolved by 2-D PAGE had coefficient of determination (R-2) values greater than 0.8, showing a linear increase in fluorescence with increasing protein concentration. The coefficient of variation (CV) was less than 50% for at least 93% of the 424 spots analyzed for both gel-to-gel variance and animal-to-animal variance. Using mitochondrial protein fractions prepared from skeletal muscle of 18-month-old mice, we show that 10 animals will be sufficient to detect a 100% difference in the 97% (i.e. 505) of the proteins resolved by 2-D PAGE. Thus, 2-D PAGE provides a sensitive and reliable technique for analysis of protein expression in mitochondria. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据