Use of cellular glucose-6-phosphate dehydrogenase for cell quantitation: applications in cytotoxicity and apoptosis assays
Use of cellular glucose-6-phosphate dehydrogenase for cell quantitation: applications in cytotoxicity and apoptosis assays
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DOI：
10.1016/j.ab.2004.02.007
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发表时间：
2004-06-01
影响因子：
2.9
通讯作者：
Zhou, MJ
中科院分区：
文献类型：
--
作者：
Batchelor, RH;Zhou, MJ
开发了基于荧光的微孔板测定，以基于葡萄糖-6-磷酸脱氢酶（G6 PD）活性的测量来定量细胞死亡。G6 PD是一种胞质酶，当质膜完整性受损时会从细胞中泄漏。在该测定中，通过偶联酶反应将细胞外G6 PD的活性与刃天青还原为荧光产物试卤灵相关联来测量细胞死亡。偶联酶反应允许在少量G6 PD前快速信号放大，这是基于单独刃天青的测定的优势。该检测方法快速、无毒，并且适合于高通量筛选。该测定具有0.78的Z'因子。(C)2004年爱思唯尔公司All rights reserved.
A fluorescence-based microplate assay was developed to quantity cell death based upon the measurement of glucose-6-phosphate dehydrogenase (G6PD) activity. G6PD is a cytosolic enzyme and leaks from cells when plasma membrane integrity is compromised. In this assay, cell death is measured by correlating the activity of extracellular G6PD to the reduction of resazurin to the fluorescent product, resorufin, via a coupled-enzyme reaction. The coupled-enzyme reaction permits rapid signal amplification front small amounts of G6PD, an advantage over assays based on resazurin alone. This assay is rapid, nontoxic, and amenable to high-through-put screening. The assay has a Z' factor of 0.78. (C) 2004 Elsevier Inc. All rights reserved.