Use of cellular glucose-6-phosphate dehydrogenase for cell quantitation: applications in cytotoxicity and apoptosis assays

被引：34

作者：

Batchelor, RH ^{[1
]}

Zhou, MJ ^{[1
]}

机构：

[1] Mol Probes Inc, Eugene, OR 97402 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2004年 / 329卷 / 01期

关键词：

fluorescence; microplate; G6PD; cell death; resorufin; resazurin; high-throughput;

D O I：

10.1016/j.ab.2004.02.007

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：35 / 42

页数：8

共 23 条

[1] A NEW RAPID AND SIMPLE NONRADIOACTIVE ASSAY TO MONITOR AND DETERMINE THE PROLIFERATION OF LYMPHOCYTES - AN ALTERNATIVE TO [H-3] THYMIDINE INCORPORATION ASSAY [J].