Caspase multiplexing:: simultaneous homogeneous time-resolved quenching assay (TruPoint) for caspases 1, 3, and 6

被引：34

作者：

Karvinen, J ^{[1
]}

Elomaa, A ^{[1
]}

Mäkinen, ML ^{[1
]}

Hakala, H ^{[1
]}

Mukkala, VM ^{[1
]}

Peuralahti, J ^{[1
]}

Hurskainen, P ^{[1
]}

Hovinen, J ^{[1
]}

Hemmilä, I ^{[1
]}

机构：

[1] PerkinElmer Life & Analyt Sci, FIN-20101 Turku, Finland

来源：

ANALYTICAL BIOCHEMISTRY | 2004年 / 325卷 / 02期

关键词：

D O I：

10.1016/j.ab.2003.10.044

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Caspases are a group of cysteine proteases involved in apoptosis and inflammation. A multiparametric homogeneous assay capable of measuring activity of three different caspases in a single well of a microtiter plate is described. Different fluorescent europium, samarium, terbium, and dysprosium chelates were coupled to a caspase substrate peptide, their luminescence properties, were analyzed, and their function in a time-resolved fluorescence quenching-based caspase 3 assay was studied. Substrates for caspases 1, 2, 3 6, and 8 and granzyme B were also synthesized and their specificities for different caspases were determined. By selecting suitable lanthanide chelates and substrates we developed a multiparametric homogeneous time-resolved fluorescence quenching-based assay for caspases 1, 3, and 6. The assay was capable of measuring the activity of both single caspases and a mixture of three caspases mixed in the same well. (C) 2003 Elsevier Inc. All rights reserved.

引用

页码：317 / 325

页数：9

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