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Determination of ATP, ADP, and AMP Levels by Reversed-Phase High-Performance Liquid Chromatography in Cultured Cells

Menegollo, M; Tessari, I; Bubacco, L; Szabadkai, G; (2019) Determination of ATP, ADP, and AMP Levels by Reversed-Phase High-Performance Liquid Chromatography in Cultured Cells. In: Raffaello, A and Vecellio Reane, D, (eds.) Calcium Signalling: Methods and Protocols. (pp. 223-232). Springer Nature: New York (NY), USA. Green open access

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Abstract

Cytoplasmic and mitochondrial Ca²⁺ signals couple cellular ATP production to activity-related energy demand. In order to accurately determine the bioenergetic effect of Ca²⁺ 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.

Type: Book chapter
Title: Determination of ATP, ADP, and AMP Levels by Reversed-Phase High-Performance Liquid Chromatography in Cultured Cells
Location: United States
ISBN-13: 978-1-4939-9017-7
Open access status: An open access version is available from UCL Discovery
DOI: 10.1007/978-1-4939-9018-4_19
Publisher version: https://doi.org/10.1007/978-1-4939-9018-4_19
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: Reversed-phase high-performance liquid chromatography (RP-HPLC), ATP/ADP ratio, Energy charge, Glycolysis, Oxidative phosphorylation, Ca²⁺ signal
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Cell and Developmental Biology
URI: https://discovery.ucl.ac.uk/id/eprint/10073232
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