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一种使用荧光染料fura-2测定无机多磷酸盐的新方法。

A novel method for determination of inorganic polyphosphates using the fluorescent dye fura-2.

作者信息

Lorenz B, Münkner J, Oliveira M P, Leitão J M, Müller W E, Schröder H C

机构信息

Institut für Physiologische Chemie, Universität, Mainz, Germany.

出版信息

Anal Biochem. 1997 Mar 15;246(2):176-84. doi: 10.1006/abio.1996.9998.

Abstract

A method for determining inorganic polyphosphate, which is based on the Mn(2+)-induced quenching of the fluorescence of the calcium indicator fura-2, is described. The effect of Mn2+ ions on fura-2 fluorescence is gradually abolished in the presence of increasing concentrations of polyphosphate; this allows the quantification both of synthetic polyphosphates and of the naturally occurring polymer isolated from tissues or cells. The described method has some advantages compared to conventional procedures for detection of polyphosphates based on the metachromatic effect on toluidine blue. It can be applied for the determination of pyrophosphate, tripolyphosphate and other short-chain polyphosphates not detectable by toluidine blue and it can be used for measurement both of pyrophosphatase and exopolyphosphatase activity.

摘要

本文描述了一种基于锰离子(Mn²⁺)诱导钙指示剂fura-2荧光猝灭来测定无机多聚磷酸盐的方法。在多聚磷酸盐浓度不断增加的情况下,Mn²⁺离子对fura-2荧光的影响会逐渐消除;这使得合成多聚磷酸盐以及从组织或细胞中分离出的天然存在的聚合物都能够被定量测定。与基于对甲苯胺蓝的异染效应检测多聚磷酸盐的传统方法相比,所描述的方法具有一些优点。它可用于测定焦磷酸盐、三聚磷酸盐和其他甲苯胺蓝无法检测到的短链多聚磷酸盐,并且可用于测量焦磷酸酶和外切多聚磷酸酶的活性。

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