![]() ![]() The data indicates that caspase 3/7 activity is detectable at 30 hours, prior to a loss of membrane integrity (CellToxTM Green) or a significant decrease in cell viability (CellTiter-Fluor™). Cell cytotoxicity, cell viability, and caspase 3/7 activity at the appropriate time point from the three separate assay readouts was normalized as a percent of the untreated controls and shown in the above graphs. Finally, 100 µL/well of Caspase-Glo® 3/7 Assay Reagent was added to all wells and luminescence was measured after a 30 minute incubation at room temperature. Then 20 µL/well of 5x CellTiter-Fluor™ reagent was added to all wells and fluorescence was measured after a 30 minute incubation at 37✬/5% CO2 in order to assess cell viability. Following the incubation periods, fluorescence was measured from the CellTox™ Green signal (cytotoxicity or loss of membrane integrity). The plates were incubated at 37✬/5% CO2 for the appropriate time (30 or 48 hours). To untreated control wells, 50 µL of medium only was added. MEGA is an integrated tool for conducting automatic and manual sequence alignment, inferring phylogenetic trees, mining web-based databases, estimating rates of molecular evolution, and testing evolutionary hypotheses. After allowing the cells to re-attach overnight at 37✬/5% CO2, 50 µL of a 2x concentrated serial titration of trastuzumab emtansine was added to the appropriate replicate wells (n = 6). Figure Legend: In two separate assay plates, 50 µL/well of HER2+ SKBR3 cells (10,000/well) were plated in medium with 2x concentrated CellTox™ Green dye. ![]()
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