4 × 105 cells per mL were seeded in 96-well plates and treated in indicated conditions (50 μL final volume). 50 μL reconstituted CaspaseGlo® 3/7 Assay substrate (Promega) were added, and the mix was incubated 30′ at room temperature. Luminescence was analysed using a Synergy HT Multi-Detection Microplate Reader (BioTek, Winooski, VT, USA) following manufacturer's recommendations.
Caspaseglo 3 7 assay substrate
Manufactured by Promega
Sourced in Germany
The CaspaseGlo® 3/7 Assay substrate is a luminescent substrate used to detect caspase-3 and caspase-7 activity in cell-based assays. It provides a straightforward method for measuring these caspase activities in a variety of experimental systems.
Automatically generated - may contain errors
3 protocols using caspaseglo 3 7 assay substrate
1
Caspase-3/7 Activation Assay
2
Caspase-Glo 3/7 Assay for Cell Viability
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Like for cell viability assays, 1×104 cells per well were seeded into a transparent 96-well microplate (CytoOne). After 24 h, the medium was replaced by serum-free medium, and cells were grown for another 24 h before treatment. After two different treatment periods of 8 and 24 h, 50 µl of Caspase-Glo 3/7 assay substrate (Promega) was added and cells were incubated for another 30 min. Then the well contents were transferred to a white-walled 96-well plate (Greiner bio-one, Germany). Luminescence was measured in a Tecan Spark multimode reader (Tecan). All treatments were measured at least in triplicates.
3
Caspase-3/7 Apoptosis Assay Protocol
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Cells were seeded like in the Resazurin assay. Five hours post‐treatment (50 µl total volume in medium −FBS), 50 µl of Caspase‐Glo 3/7 assay substrate (Promega, Mannheim, Germany) were added and the assay performed according to the manufacturer's instructions. Luminescence was measured in a Spark multimode reader (Tecan). Independent experiments were measured in duplicates.
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