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 proteases may initiate or propagate proapoptotic signals. Cathepsins are lysosomal enzymes that are also used as sensitive markers in various toxicological investigations. The Cathepsin-H Activity Assay kit is a fluorescence-based assay that utilizes the preferred cathepsin-H substrate Arginine labeled with AFC (amino-4-trifluoromethyl coumarin). Cell lysates or other samples that contain cathepsin-H will cleave the synthetic substrate R-AFC to release free AFC. The released AFC can easily be quantified using a fluorometer or fluorescence plate reader. The cathepsin-H assay is simple, straightforward, and can be adapted to 96-well plate assays. Assay conditions have been optimized to obtain the maximal activity. Sample types: cell lysates, tissue homogenates
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This assay provides for the detection of Cathepsin-H in various biological sample types. Detects alternative proteolytic enzymes such as the lysosomal cathepsin proteases that can initiate or propagate proapoptotic signals.