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Lyophilized ATP

Lyophilized ATP

公司名称: Roche Diagnostics
产品编号: 11699695001
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Intracellular Assessment of ATP Levels in Caenorhabditis elegans
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Date:
2016-12-05
[Abstract]  Eukaryotic cells heavily depend on adenosine triphosphate (ATP) generated by oxidative phosphorylation (OXPHOS) within mitochondria. ATP is the major energy currency molecule, which fuels cell to carry out numerous processes, including growth, differentiation, transportation and cell death among others (Khakh and Burnstock, 2009). Therefore, ATP levels can serve as a metabolic gauge for cellular homeostasis and survival (Artal-Sanz and Tavernarakis, 2009; Gomes et al., 2011; Palikaras et al., 2015). In this protocol, we describe a method for the determination of ... [摘要]  真核细胞严重依赖于由线粒体内的氧化磷酸化(OXPHOS)产生的三磷酸腺苷(ATP)。 ATP是主要能量货币分子,其促进细胞进行许多过程,包括生长,分化,运输和细胞死亡等(Khakh和Burnstock,2009)。因此,ATP水平可以作为细胞内稳态和存活的代谢指标(Artal-Sanz和Tavernarakis,2009; Gomes等人,2011; Palikaras等人, 2015)。在本议定书中,我们描述了使用生物发光方法在线虫秀丽隐杆线虫中测定细胞内ATP水平的方法。
关键词:老化,ATP,线粒体,能量稳态,荧光素酶,代谢,线粒体

[背景] 线粒体衍生的ATP在多种细胞和代谢中起着至关重要的作用过程。因此,已经开发了几种方法来测量这种重要代谢物的水平(Drew和Leeuwenburgh,2003; Khan,2003; Lagido等人,2001; Vives-Bauza等人, ,2007)。在1947年,McElroy提出使用萤火虫生物发光来确定细胞内ATP水平,当他发现ATP在光生产中的关键作用(McElroy,1947)。克隆和重组蛋白质技术的发展促进了萤火虫荧光素酶测定法的开发以测定ATP水平(de Wet等人,1985)。萤火虫荧光素酶,单体61 ...

Determination of the H+-ATP Synthase and Hydrolytic Activities
Author:
Date:
2016-08-20
[Abstract]  The H+-ATP synthase of the inner mitochondrial membrane utilizes the proton gradient generated by the respiratory chain to synthesize ATP. Under depolarizing conditions, it can function in reverse by hydrolyzing ATP to generate a proton gradient. The protocols presented here allow the facile determination of both the synthetic and hydrolytic activities of the H+-ATP synthase in isolated mitochondria and in permeabilized mammalian cells. Since the protocol requires the isolation of polarized and well-coupled mitochondria, first we describe the protocol for mitochondrial ... [摘要]  内线粒体膜的H sup + -ATP合酶利用呼吸链产生的质子梯度来合成ATP。 在去极化条件下,它可以通过水解ATP产生质子梯度而反向作用。 本文提供的方案允许容易地测定分离的线粒体和透化的哺乳动物细胞中H sup + -ATP合酶的合成和水解活性。 由于该协议需要极化和良好耦合的线粒体的分离,首先我们描述线粒体从小鼠组织分离的协议。 第二,我们描述了用于测量ATP合成活性作为终点和在分离的线粒体和透化细胞中的动力学模式的方案。 最后,我们描述了用于测定酶在分离的线粒体中的ATP水解活性的方案。

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