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培养的哺乳动物细胞中5',5'''-P1,P4-四磷酸二腺苷水平的测定

Determination of diadenosine 5',5''',-P1,P4-tetraphosphate levels in cultured mammalian cells.

作者信息

Baker J C, Jacobson M K

出版信息

Anal Biochem. 1984 Sep;141(2):451-60. doi: 10.1016/0003-2697(84)90070-8.

DOI:10.1016/0003-2697(84)90070-8
PMID:6093629
Abstract

A simple method for measuring the cellular content of diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) in cultured mammalian cells is described. Ap4A was rapidly extracted by dissolving cell monolayers using 0.1 N NaOH. It was separated from the bulk of cellular components in a single step by selective adsorption to a highly specific boronate affinity resin. Subpicomole amounts were quantified by a luciferin-luciferase bioluminescence assay performed in the presence of alkaline phosphatase and venom phosphodiesterase. The selectivity of this assay for Ap4A in cultured mouse cells was established by high-performance liquid chromatography. This method allows the routine measurement of subpicomole amounts of Ap4A derived from a single dish of cells.

摘要

本文描述了一种用于测量培养的哺乳动物细胞中二腺苷 5',5'''-P1,P4-四磷酸（Ap4A）细胞含量的简单方法。通过使用 0.1 N NaOH 溶解细胞单层快速提取 Ap4A。通过选择性吸附到高度特异性的硼酸酯亲和树脂上，在一步中将其与大部分细胞成分分离。通过在碱性磷酸酶和蛇毒磷酸二酯酶存在下进行的荧光素-荧光素酶生物发光测定法定量亚皮摩尔量。通过高效液相色谱法确定了该测定法对培养的小鼠细胞中 Ap4A 的选择性。该方法允许常规测量来自单个培养皿细胞的亚皮摩尔量的 Ap4A。

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Determination of diadenosine 5',5''',-P1,P4-tetraphosphate levels in cultured mammalian cells.培养的哺乳动物细胞中5',5'''-P1,P4-四磷酸二腺苷水平的测定
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引用本文的文献

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Mol Cell Biol. 1986 Apr;6(4):1179-86. doi: 10.1128/mcb.6.4.1179-1186.1986.
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Intracellular 5',5'-dinucleoside polyphosphate levels remain constant during the Escherichia coli cell cycle.在大肠杆菌细胞周期中，细胞内5',5'-二核苷多磷酸水平保持恒定。
J Bacteriol. 1987 Jan;169(1):419-22. doi: 10.1128/jb.169.1.419-422.1987.
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Cell cycle variations of dinucleoside polyphosphates in synchronized cultures of mammalian cells.
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