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小鼠脑内ATP的生物发光测定:提高检测灵敏度的决定因素

Bioluminometric assay of ATP in mouse brain: Determinant factors for enhanced test sensitivity.

作者信息

Khan Haseeb Ahmad

机构信息

Department of Medical Research, Riyadh Al Kharj Hospital Program, Riyadh 11159, Kingdom of Saudi Arabia.

出版信息

J Biosci. 2003 Jun;28(4):379-82. doi: 10.1007/BF02705114.

Abstract

Firefly luciferase bioluminescence (FLB) is a highly sensitive and specific method for the analysis of adenosine-5-triphosphate (ATP) in biological samples. Earlier attempts to modify the FLB test for enhanced sensitivity have been typically based on in vitro cell systems. This study reports an optimized FLB procedure for the analysis of ATP in small tissue samples. The results showed that the sensitivity of the FLB test can be enhanced several fold by using ultraturax homogenizer, perchloric acid extraction, neutralization of acid extract and its optimal dilution, before performing the assay reaction.

摘要

萤火虫荧光素酶生物发光法(FLB)是一种用于分析生物样品中三磷酸腺苷(ATP)的高灵敏度和特异性方法。早期为提高灵敏度而对FLB测试进行改进的尝试通常基于体外细胞系统。本研究报告了一种用于分析小组织样品中ATP的优化FLB程序。结果表明,在进行测定反应之前,通过使用超声均质器、高氯酸提取、酸提取物的中和及其最佳稀释,可以将FLB测试的灵敏度提高几倍。

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