Luminescence (RLU) Change in Cell Viability Compared to Vehicle Control (fold) 12466MA TIPS & TECHNIQUES Real-Time Monitoring of Cell Viability Using a Plate-Reader T he RealTime-Glo™ Assay enables monitoring of cell viability continuously in real time for up to 72 hours from the same sample well. This non-toxic, non-lytic assay provides an easy way to collect information on viable cell number at any time during the course of experiments, and allows you to respond immediately to changes in viability by performing a follow-up assay or molecular application. By generating multiple data points from a single well, it also saves time, reagents, compounds, plates and labor. Cell Viability Changes Monitored for up to 72h in a Single Plate Figure 1 shows dose-response curves demonstrating the effect of bortezomib on A549 cells using the RealTime-Glo™ Assay. The assay reagents were added to the cells at plating and viability monitored every hour for 72 hours from each well. The vehicle control shows how luminescence increases as the untreated cells proliferate, and decreases as the dose of bortezomib increases and the cells begin to die. Both NanoLuc® luciferase and the prosubstrate are extremely stable under cell culture conditions, allowing monitoring of cell viability over an extended incubation period. A. 100,000 150,000 200,000 250,000 300,000 50,000 0 020406080 Time (hours) 0.0 –1 01 23 Bortezomib (log nM) 12h EC50 Value (nM) 3.07 24h 2.29 Incubation Time 36h 48h 2.27 1.99 60h 1.78 72h 1.62 Figure 1. EC50 determination using the continuous-read assay format. Five hundred A549 cells/well were plated in a 384-well plate in 40µl of cell culture medium containing 2X RealTime-Glo™ reagent. An equal volume of 2X bortezomib was added to achieve the indicated concentrations. Panel A. Luminescence (cell viability) was monitored every hour for 72 hours. Panel B. The fold change in cell viability of bortezomib-treated cells compared to the vehicle control at each concentration. enews | September 2015 300nM 150nM 75nM 37.5nM 18.75nM 9.38nM 4.69nM 2.34nM 1.17nM 0.59nM 0.29nM 0nM B. 1.5 12h 48h 1.0 24h 60h 36h 72h 0.5 Promotion
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