Nanosecond time-resolved fluorescence protease assays

被引：25

作者：

Hennig, Andreas

Roth, Doris

Enderle, Thilo

Nau, Werner M.

机构：

[1] Int Jacobs Univ Bremen, Sch Engn & Sci, D-28759 Bremen, Germany

[2] F Hoffmann La Roche & Cie AG, CH-4070 Basel, Switzerland

来源：

CHEMBIOCHEM | 2006年 / 7卷 / 05期

关键词：

biosensors; enzymes; fluorescence; proteases; time-resolved assays;

D O I：

10.1002/cbic.200500561

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

A background reduction of several orders of magnitude in fluorescence protease assays can be achieved by employing time-resolved fluorescence detection with a time gate to eliminate short-lived emission. The use of 2,3-diazabicyclo[2.2.2]oct-2-ene as a hydrophilic and biocompatible fluorophore allows such measurements to be conducted in the nanosecond time domain with a purely organic fluorophore for the first time. The method is applicable to assaying carboxypeptidase A activity on both the laboratory scale and in high-throughput screening. © 2006 Wiley-VCH Verlag GmbH & Co. KGaA.

引用

收藏

页码：733 / 737

页数：5

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