Product Handling - Assay Method (NBP2-29396):
Materials:
Dissolve 1mg of Z-D(OMe)E(OMe)VD(OMe)-FMK in DMSO to get appropriate concentration:
75 ul DMSO = 20mM
150 ul DMSO = 10 mM
300 ul DMSO = 5 mM
Method:
Add 2 ul of above stock solutions to 1 ml of culture medium containing cells to give final DMSO concentration of
0.2%. Levels of DMSO above this may cause some cellular toxicity thus masking the effect of the ICE-protease
inhibitors. Two microliter of 10mM stock solution in 1 ml medium = 20 uM final Z-D(OMe)E(OMe)VD(OMe)-FMK
concentration. Caspase inhibitors have been successfully used in tissue cultures to inhibit apoptosis at final working
concentrations of approximately 50 nM to 100 uM. The variability reflects cell type and method. Investigators should
determine the optimal concentration empirically.
