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Characteristics
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Peptide-fluoromethyl ketone inhibitor of caspases. BOC-D-FMK is a pan-caspase inhibitor. The -CH2F (fluoromethyl ketone) inhibitor has several advantages over other types of derivatives: Penetrates cell membranes, Not toxic to cells, Irreversible inhibition. Binds irreversibly with and inhibits Caspase-1/ICE and Caspase-4.
Dissolve the BOC-D-FMK Inhibitor in DMSO before use.
For use on intact cells:
1. Prepare desired concentrated stock solutions as follows:
1 mg BOC-D(OMe)-FMK
in 19μl DMSO = 20mM
in 38μl DMSO = 10 mM
in 76μl 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 0.2% may cause some cellular toxicity, thus masking the effect of the BOC-D-FMK protease inhibitor. Adding 2 μL of a 10 mM stock solution to 1 mL of culture medium gives a final BOC-D- FMK concentration of 20 mM. Typical final concentrations are 5-20 mM.
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.
IMPORTANT NOTE for in vitro use: Our peptide inhibitors are synthesized as methyl esters to enhance cell permeability. In intact cells, the methyl groups are removed by endogenous enzymes. For in vitro experiments with purified enzymes, however, the methyl groups must first be removed by treating the inhibitor with esterase. A procedure is available upon request.
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