Chida Junji, Kido Hiroshi
Division of Enzyme Chemistry, Institute for Enzyme Research, The University of Tokushima, Tokushima, Japan.
Methods Mol Biol. 2014;1098:21-32. doi: 10.1007/978-1-62703-718-1_2.
Adenosine 5'-triphosphate (ATP) is the "energy currency" of organisms and plays central roles in bioenergetics, whereby its level is used to evaluate cell viability, proliferation, death, and energy transmission. In this chapter, we describe an improved and efficient method for extraction of ATP from tissues and cells using phenol-based reagents. The chaotropic extraction reagents reported so far co-precipitate ATP with insoluble proteins during extraction and with salts during neutralization. In comparison, the phenol-based reagents extract ATP well without the risks of co-precipitation. The extracted ATP can be quantified by the luciferase assay or high-performance liquid chromatography.
腺苷 5'-三磷酸（ATP）是生物体的“能量货币”，在生物能量学中发挥核心作用，其水平用于评估细胞活力、增殖、死亡和能量传递。在本章中，我们描述了一种使用酚类试剂从组织和细胞中提取 ATP 的改进高效方法。迄今为止报道的离液剂提取试剂在提取过程中会使 ATP 与不溶性蛋白质共沉淀，在中和过程中会与盐共沉淀。相比之下，酚类试剂能很好地提取 ATP，且不存在共沉淀风险。提取的 ATP 可通过荧光素酶测定法或高效液相色谱法定量。
