Caspase 3/7 activity assay

MG Maria Jesús García-Murria
GD Gerard Duart
BG Brayan Grau
ED Elisabet Diaz-Beneitez
DR Dolores Rodríguez
IM Ismael Mingarro
LM Luis Martínez-Gil
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Quantification of caspase activity was carried out by using the Caspase-Glo 3/7 assay kit (Promega) following the protocol recommended by the supplier. HeLa cell monolayers grown in 24-well plates were infected and transfected, in duplicate, under the conditions indicated above. At 24 h.p.i. cells were harvested in medium and kept frozen until their analysis. 25 µL of the cell lysates under study was added to 25 µL of Caspase-Glo 3/7 reagent in a 96-well plate. Plates were gently shaken and then incubated in the dark at room temperature for 60 min before recording the luciferase activity by using an Appliskan luminometer (ThermoScientific).

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