A Real-Time Cytotoxicity Assay That Delivers More Relevant Data
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Andrew L. Niles is a Senior Research Scientist in the Assay Designs Group at Promega Corporation (Madison, WI). His laboratory is focused on developing homogeneous :add-mix-measure" assay chemistries for cell-based studies. The lab utilizes fluorescent and/or bioluminescent detection platforms configured in plate-based, multiplexed formats to detect cytotoxicity and elucidate mechanism of action. These efforts have led to multiple commercialized assay systems in the area of in vitro toxicity testing. He has been awarded eleven U.S. Patents relating to cell and protease biology, authored several book chapters and reviews, and is an active member of Society of Toxicology, Society for Laboratory Automation, Laboratory Robotics Interest Group, and American Association for Cancer Research.
Andrew L. Niles
Sr. Research Scientist
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- Original Webinar Date: Tuesday, September 9, 2014
This webinar describes how a new, real-time assay can be conveniently employed to reveal the kinetics and magnitude of response for any individual cytotoxic profile without the limitations of conventional approaches. When combined with value-added multiplex chemistries, these data can provide important information for defining mechanism of action.
Cytotoxicity assay chemistries typically measure nascent biomarkers which emerge from cultures of cells as a result of cell death. These gain-of-signal endpoint assays typically rely on biomarkers which have enzymatic activities, and are therefore subject to time-dependent biomarker degradation. Because the rate of emergence (kinetics) of these cytotoxicity biomarkers vary greatly for any individual cytotoxic response, conventional cytotoxicity assays are best employed as parallel endpoints in time course measures. In an effort to address this duplication of effort and inefficiency, we have developed a physiologically inert, pro-fluorescent cytotoxicity assay probe which can be delivered to cultures at the time of dosing and be used as a real time measure of the cytotoxic phenotype. The probe is not subject to signal degradation and serves as an accumulative measure of cytotoxicity in cell cultures out to 72hrs or more of exposure. Importantly, the probe can be used to define the beginnings of cytotoxic stress, allowing for same-well multiplexing with spectrally distinct assays which help elucidate mechanism of action. This webinar will showcase the utility of the probe in defining specific effects from multiple compounds on diverse cell types.