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Comment
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Fluoromethyl ketone peptide inhibitor of cathepsin B (caspase inhibitor negative control). The CH2F (fluoromethyl ketone) inhibitor has several advantages over other types of derivatives: Penetrates cell membranes, is nontoxic to cells, irreversible inhibition.
The caspases are involved in the proteolytic cascade that results in apoptosis and in the maturation process of IL-1? necessary for inflammation. The increasing number of FMKtype inhibitors for various caspases require an FMK-type inhibitor that does not affect caspases as a negative control. The Cathepsin B Inhibitor does not inhibit the activity of caspases in intact cells and does not block the induction of apoptosis as caspase inhibitors do. It can, therefore, be used as a negative control for the FMK moiety of any caspase inhibitor, both in vivo and in vitro.
Specifically binds to and inhibits various cysteine proteases (those not requiring P1 Asp) such as cathepsin B. Does not bind to or inhibit any of the caspases.
Dissolve Cathepsin B Inhibitor in high purity (>99.9%) DMSO before use.
For use on intact cells:
1. Prepare concentrated stock solution in DMSO as follows:
Dissolve 5 mg Z-FA-FMK
in 1,295 μl DMSO = 10 mM 2. Add 2 μL of the 10 mM stock solution to 1 mL of culture medium gives a 20 μM final Z-FA- FMK concentration.
A. Inhibitor of cysteine proteases (non-ICE (caspase) proteases): Irreversible inhibition of various cysteine proteases at an effective concentration of approximately 0.5-10 μM.
B. Use as a Negative Control for ICE- (caspase) proteases: Effective final concentrations of the Caspase FMK Inhibitors are estimated to be 5-20 μM. Use of Z-FA- FMK as a Negative Control Inhibitor should be in the same range.
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