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|Title:||A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents||Authors:||Zhu, Xiaoming
Luo, Kathy Qian
|Issue Date:||2012||Source:||Zhu, X., Fu, A., & Luo, K. Q. (2012). A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents. Biochemical and Biophysical Research Communications, 418(4), 641-646.||Series/Report no.:||Biochemical and biophysical research communications||Abstract:||In this study, we developed a high-throughput endothelial cell apoptosis assay using a fluorescence resonance energy transfer (FRET)-based biosensor. After exposure to apoptotic inducer UV-irradiation or anticancer drugs such as paclitaxel, the fluorescence of the cells changed from green to blue. We developed this method into a high-throughput assay in 96-well plates by measuring the emission ratio of yellow fluorescent protein (YFP) to cyan fluorescent protein (CFP) to monitor the activation of a key protease, caspase-3, during apoptosis. The Z′ factor for this assay was above 0.5 which indicates that this assay is suitable for a high-throughput analysis. Finally, we applied this functional high-throughput assay for screening vascular disrupting agents (VDA) which could induce endothelial cell apoptosis from our in-house compounds library and dioscin was identified as a hit. As this assay allows real time and sensitive detection of cell apoptosis, it will be a useful tool for monitoring endothelial cell apoptosis in living cell situation and for identifying new VDA candidates via a high-throughput screening.||URI:||https://hdl.handle.net/10356/100445
|ISSN:||0006-291X||DOI:||http://dx.doi.org/10.1016/j.bbrc.2012.01.066||Rights:||© 2013 Elsevier Inc.||Fulltext Permission:||none||Fulltext Availability:||No Fulltext|
|Appears in Collections:||SCBE Journal Articles|
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