Comment
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Peptide-fluoromethyl ketone inhibitor of caspase-8(FLICE). The CH2F or FMK (fluoromethyl ketone) inhibitor has several advantages over other types of derivatives: Penetrates cell membranes, is nontoxic to cells, irreversible inhibition. Caspase-8 (also known as FLICE, MACH or Mch5) is a member of the caspase family of cysteine proteases. Caspases play an important role in apoptosis signaling and effector mechanisms. Caspase-8 is most similar to caspase-10, both of which have "death domain" motifs. Caspase-8 appears to be physically associated with the signaling mechanism during Fas-mediated cell death and its association with Fas or tumor necrosis factor receptors via an interaction with FADD suggests it functions as an initiator rather than an effector of the cell death pathway. Thus, caspase-8 is an upstream activator in the protease cascade that proteolytically matures other caspases. Highly specific inhibitor of caspase-8. May have some inhibitory activity towards caspase-6. For caspase-8 fluorometric assays using the Caspase-8(FLICE) Fluorogenic Substrate, AcIETD-AFC (ABIN924941), Caspase-8(FLICE) Inhibitor can be used to assess the contribution of contaminating proteases to the overall rate of proteolysis. Dissolve the Caspase-8(FLICE) Inhibitor in high purity (>99.9%) DMSO before use.
For use on intact cells: 1. Prepare desired concentrated stock solutions as follows: 3 mg Z-IETD-FMK in 45μl DMSO = 10 mM
2. Adding 2 μL of a 10 mM stock solution to 1 mL of culture medium gives a final Z-IETD- FMK concentration of 20 μM. Effective final concentrations are estimated to be 5-20 μM.
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