The serine/threonine AMP-activated protein kinase (AMPK) is a central player in the regulation of energy homeostasis, and its activity is tightly controlled, among other mechanisms, by subtle changes in cellular adenine nucleotide levels. In this chapter, we describe a step-by-step protocol for rapid, highly sensitive, reproducible, and simultaneous determination of ATP, ADP, and AMP concentrations in cell or tissue samples by reversed-phase high-performance liquid chromatography (HPLC).
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