Cells were treated GST-TRAIL or drozibumab as indicated in the text and figure legends. For caspase inhibition, cells were preincubated with 100 uM ZVAD-FMK or DMSO for two hours and subsequently incubated overnight with 2500 ng/ml drozitumab or 2500 ng/ml drozitumab plus 10 μg/ml F(ab’)2. Caspase activity was assessed using the Caspase-Glo® 3/7 assay system (Cat # G8092, Promega Corporation, Madison, WI) and/or Caspase-Glo® 8 assay system (Cat # G8202, Promega Corporation, Madison, WI) as previously described [27 (link)]. Three independent experiments normalized to the control cells +/− SE were performed.
Protocol detail
Caspase Activation Assay for TRAIL
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Corresponding Organization :
Other organizations : National Cancer Institute, National Institutes of Health, Center for Cancer Research, MedStar Washington Hospital Center, University of Missouri
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Variable analysis
independent variables
- GST-TRAIL treatment
- Drozibumab treatment
- ZVAD-FMK preincubation
- Drozitumab (2500 ng/ml) treatment
- Drozitumab (2500 ng/ml) plus F(ab')2 (10 μg/ml) treatment
- Caspase activity assessed using the Caspase-Glo® 3/7 assay system
- Caspase activity assessed using the Caspase-Glo® 8 assay system
- DMSO preincubation
- Control cells
- Positive control: Not specified
- Negative control: Control cells
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