What is the optimal time point to measure Caspase activity after Z-VAD-FMK treatment?

The window is narrow and induction-dependent:

• Pre-incubation: Always pre-treat cells with Z-VAD-FMK for 30–60 minutes before adding the apoptosis inducer.
• Early Detection: For Caspase activity assays, the peak signal usually occurs 2–6 hours post-induction in the presence of Z-VAD-FMK.
• Stability: If you wait too long (e.g., 24 hours), Z-VAD-FMK might lose its potency due to degradation, allowing Caspases to reactivate.