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Comment
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Lyophilized solid. Peptide-fluoromethyl ketone inhibitor of caspases.
The CH2F (fluoromethyl ketone) inhibitor has several advantages over other types of derivatives: Penetrates cell membranes, Not toxic to cells, Irreversible inhibition
Available data indicates that FMK inhibitors based on the sequence AEVD will inactivate caspases-6, 8, 9, and to a lesser extent caspases-1 and 3. Caspases-2, 4, 5, 7, and 10 will be little affected.
Dissolve the Caspase-13 Inhibitor in DMSO before use. It is important that dry, good quality DMSO be used.
For use on intact cells:
1. Prepare desired concentrated stock solutions as follows:
3 mg Z-AEVD-FMK
in 24μl DMSO = 20 mM
in 49μl DMSO = 10 mM
in 98μl DMSO = 5 mM, etc.
2. Add 2 μl of above stock solution to 1 ml culture medium containing cells such that the fμl DMSO concentration is 0.2%. Levels of DMSO above this may be toxic, thus masking the effect of the Caspase-13 protease inhibitor. Adding 2 μl of a 10 mM stock solution to 1 ml of culture medium gives a final Z-AEVD-FMK concentration of 20 μM. Typical final concentrations are 5 - 20 μM.
For extended use in vivo and in vitro:
For experiments extending 12 to 48 hours, fresh inhibitor may have to be added (injected) due to inactivation of the inhibitor by endogenous cysteine proteases.
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