利用反相高效液相色谱法测定培养细胞中的ATP、ADP和AMP水平。
Determination of ATP, ADP, and AMP Levels by Reversed-Phase High-Performance Liquid Chromatography in Cultured Cells.
作者信息
Menegollo Michela, Tessari Isabella, Bubacco Luigi, Szabadkai Gyorgy
机构信息
Department of Biomedical Sciences, University of Padua, Padua, Italy.
Department of Biology, University of Padua, Padua, Italy.
出版信息
Methods Mol Biol. 2019;1925:223-232. doi: 10.1007/978-1-4939-9018-4_19.
Cytoplasmic and mitochondrial Ca signals couple cellular ATP production to activity-related energy demand. In order to accurately determine the bioenergetic effect of Ca signals, cellular energy charge, i.e., the compound ratio of the phosphorylated adenine nucleotides AMP, ADP, and ATP, should be estimated. Reversed-phase high-performance liquid chromatography (RP-HPLC) allows the rapid separation and quantitation of these molecules. Here we describe a protocol applied in our laboratories to quantify ATP, ADP, and AMP nucleotides in cellular extracts.
细胞质和线粒体中的钙信号将细胞ATP生成与活动相关的能量需求联系起来。为了准确确定钙信号的生物能量效应，应估算细胞能量电荷，即磷酸化腺嘌呤核苷酸AMP、ADP和ATP的复合比率。反相高效液相色谱法（RP-HPLC）能够快速分离和定量这些分子。在此，我们描述一种在我们实验室中用于定量细胞提取物中ATP、ADP和AMP核苷酸的方法。
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