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  Adenine Nucleotide and Nicotinamide Adenine Dinucleotide Measurements in Plants

Zhang, Y., Krahnert, I., Bolze, A., Gibon, Y., & Fernie, A. R. (2020). Adenine Nucleotide and Nicotinamide Adenine Dinucleotide Measurements in Plants. Current Protocols in Plant Biology, 5(3): e20115. doi:10.1002/cppb.20115.

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Zhang, YJ1, Autor           
Krahnert, I.1, Autor           
Bolze, A.1, Autor           
Gibon, Yves2, Autor
Fernie, A. R.1, Autor           
Affiliations:
1Central Metabolism, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society, ou_1753339              
2External Organizations, ou_persistent22              

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Schlagwörter: ADP, ATP, NAD+/NADP+, NADH/NADPH
 Zusammenfassung: Abstract As the principal co-factors of many metabolic pathways, the measurement of both adenine nucleotides and nicotinamide adenine dinucleotide provides important information about cellular energy metabolism. However, given their rapid and reversible conversion as well as their relatively low concentration ranges, it is difficult to measure these compounds. Here, we describe a highly sensitive and selective ion-pairing HPLC method with fluorescence detection to quantify adenine nucleotides in plants. In addition, nicotinamide adenine dinucleotide is a crucially important redox-active substrate for multiple catabolic and anabolic reactions with the ratios of NAD+/NADH and NADP+/NADPH being suggested as indicators of the general intracellular redox potential and hence metabolic state. Here, we describe highly sensitive enzyme cycling−based colorimetric assays (with a detection limit in the pmol range) performed subsequent to a simple extraction procedure involving acid or base extraction to allow the measurement of the cellular levels of these metabolites. © 2020 The Authors. Basic Protocol 1: Preparation of plant material for the measurement Basic Protocol 2: Measurement of ATP, ADP, and AMP via HPLC Basic Protocol 3: NAD+/NADP+ measurements Basic Protocol 4: NADH/NADPH measurements Basic Protocol 5: Data analysis and quality control approaches

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Sprache(n): eng - English
 Datum: 2020
 Publikationsstatus: Erschienen
 Seiten: -
 Ort, Verlag, Ausgabe: -
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 Identifikatoren: DOI: 10.1002/cppb.20115
BibTex Citekey: doi:10.1002/cppb.20115
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Titel: Current Protocols in Plant Biology
  Kurztitel : Curr Protoc Plant Biol
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: Chichester : Wiley
Seiten: - Band / Heft: 5 (3) Artikelnummer: e20115 Start- / Endseite: - Identifikator: ISSN: 2379-8068
CoNE: https://pure.mpg.de/cone/journals/resource/2379-8068