文章基本信息

标题：Multiplexing fluorogenic esterase-based viability assay with luciferase assays
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作者：Kenji Ohgane ; Hiromasa Yoshioka ; Yuichi Hashimoto 等
期刊名称：MethodsX
印刷版ISSN：2215-0161
电子版ISSN：2215-0161
出版年度：2019
卷号：6
页码：2013-2020
DOI：10.1016/j.mex.2019.09.008
语种：English
出版社：Elsevier
摘要：Graphical abstractDisplay OmittedAbstractLuciferase-based reporter assays are one of the most common cell-based screening formats for drug discovery, and simultaneous evaluation of the cytotoxic effect of test compounds is of great value in reducing false-positives. Here we share a multiplex assay protocol that allows sequential measurement of cell viability (cell number) and luciferase activity of the same sample in a multi-well-plate format. The viability assay employs a fluorogenic esterase substrate, CytoRed.•This protocol allows sequential measurement of endogenous esterase activity (as a surrogate for cell number) and then luciferase activity in a single sample.•The protocol eliminates the need for parallel viability assay or protein assay using separate aliquots of the lysate.•This protocol is especially useful for assays with cells stably expressing a luciferase construct, for which co-transfection of another reporter gene is not a viable option.
关键词：Multiplex assay;Luciferase;CytoRed;Esterase;Fluorogenic substrate;Viability