What is the optimal Puromycin concentration that should be used when selecting AML12 cells?

12 September 2018 0 9K Report

I am performing a kill curve with puromycin, ranging from 0.5 ug/ml to 10 ug/ml. It has been 6 days from initial treatment and 100% of the cells have not been killed as yet. Cells have died but there cells still attached to the plate. Should I split the cells to ensure that these cells are in fact dying (should not re-attach)? The cells are clustered and resembles that of unhealthy or dying cells, yet they remain attached. What is the typical puromycin concentration and time course for effective selection? Also what do you suggest to resolve this issue?

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