A Colorimetric Assay of Citrate Synthase Activity in <em>Drosophila Melanogaster</em>

Ping Wei; Qihong Liu; Wen Xue; Jiwu Wang

doi:10.3791/59454

JoVE Journal
Biochemistry

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JoVE Journal Biochemistry

A Colorimetric Assay of Citrate Synthase Activity in Drosophila Melanogaster

果蝇梅拉诺加斯特的锡酸盐合成酶活性的色度测定

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04:30 min

January 16, 2020

DOI: 10.3791/59454-v

Ping Wei*¹, Qihong Liu*², Wen Xue², Jiwu Wang²

¹Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai Clinical Center for Diabetes,Shanghai Jiao Tong University Affiliated Sixth People's Hospital, ²Department of Anatomy of Physiology,Shanghai Jiao Tong University School of Medicine

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我们提出了一种对胆酸合成酶活性的色度测定方案，用于定量果蝇组织均质中完整的线粒体质量。

果蝇是研究线粒体功能的好模型系统。在这里，我们提出了一个协议，用于测量成人苍蝇组织中柠檬酸盐合成酶活性。该协议不需要线粒体隔离。

它快速而简单，适合测量幼虫、培养细胞和哺乳动物组织中柠檬酸盐合成酶活性。首先收集每个样本的成年苍蝇，确保收集每个基因型至少三分样本。在每个样品的 1.5 毫升试管中制备 500 微升的冰冷萃取缓冲液，内有 20 毫摩尔 HEPES、1 毫摩尔 EDTA 和 0.1%三吨 X-100。

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生物化学问题 155 线粒体质量锡酸盐合成酶活性色度测定果蝇黑色素测定线粒体功能障碍代谢研究