Evaluation of Mitochondrial Function v2

Evaluation of Mitochondrial Function v2
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线粒体功能评估 v2

DOI:
10.17504/protocols.io.7s8hnhw
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发表时间:
2019
期刊:
protocols.io
影响因子:
--
通讯作者:
E. Abel
E. Abel
中科院分区:
--
文献类型:
--
作者:
E. Abel

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本协议描述了DiaComp用于评估线粒体功能的程序。从线粒体基质中释放柠檬酸合成酶,稀释为1:10。反应在1ml反应缓冲液中进行,该反应缓冲液含有20 HEPES,EGTA,220蔗糖,40 KCl,0.1 DTNB,0.1乙酰辅酶A,25°C,pH 7.4。反应开始加入0.05 mM草酰乙酸酯(基线省略),在412 nm处用超声波3000分光光度计监测3分钟。超声波
This protocol describes the procedures used by the DiaComp for evaluating mitochondrial function. liberate citrate synthase from mitochondrial matrix and diluted 1:10. reaction is then performed in 1 ml reaction buffer containing 20 HEPES, EGTA, 220 sucrose, 40 KCl, 0.1 DTNB, and 0.1 acetyl-CoA, pH 7.4 at 25°C. The reaction is started by adding 0.05 mM oxaloacetate (omitted at baseline) and monitored at 412 nm for 3 min using an Ultraspec 3000 spectrophotometer. Ultraspec