Apoptosis can
be mediated by mechanisms other than the traditional caspase-mediated
cleavage cascade. There is growing recognition that
alternative proteolytic enzymes such as the lysosomal
cathepsin protease may intiate or propagate proapoptotic
signals, but it is currently unclear how cathepsin mediate
apoptosis. Cathepsin are lysosomal enzymes that are also
used as sensitive markers in various toxicological
investigations. BioVision's newly developed Cathepsin
Activity Assay kits are fluorescence-based assays that
utilize the preferred substrate sequence for each cathepsin,
labeled with AFC (amino-4-trifluoromethyl coumarin). Cell
lysates or other samples that contain cathepsins will cleave
the synthetic substrate to release free AFC. The released
free AFC can easily be quantified using a fluorometer of
fluorescence plate reaser. The cathepsin assays are simple,
straightforward, and can be adapted to 96-well plate assays.
Assay conditons have been optimized to obtain the maximal
activity. Active cathepsin and cathepsin inhibitors are also
available.
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