Hi all,

I am using a MUSE caspase 3/7 kit for flow that requires a positive control for apoptosis. I am studying fibroblast apoptosis. I UV-irradiated fibroblasts with a portable UV box for 5 minutes and tested them 16 hours later for the positive control. I used this data to calibrate the machine.

Fibroblasts treated with H2O2 for 48 hours at 300 micromolar showed increased apoptosis over controls, but control cells also showed apoptosis at an unexpectedly high rate. I think my positive control for machine calibration was not very good. Can anybody provide some suggestions how to prepare a good positive control that I can try? Thanks for your time.

Mel