Product Overview :
The cathepsin-K assay is simple, straightforward, and can be adapted to 96-well plate assays. Assay conditions have been optimized to obtain the maximal activity.
Description :
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-K Activity Assay kit is a fluorescence-based assay that utilizes the preferred cathepsin-K substrate sequence LR labeled with AFC (amino-4-trifluoromethyl coumarin). Cell lysates or other samples that contain cathepsin-K will cleave the synthetic substrate LR-AFC to release free AFC. The released AFC can easily be quantified using a fluorometer or fluorescence plate reader.
Applications :
Detects alternative proteolytic enzymes such as the lysosomal cathepsin proteases that can initiate or propagate proapoptotic signals.
Stability :
Store kit at -20°C (Store CK Cell Lysis Buffer and CK Reaction Buffer at 4°C after opening). Protect CK Substrate from light. All reagents are stable for 6 months under
proper storage conditions.
Kit Components :
CK Cell Lysis Buffer 25 ml
CK Reaction Buffer 5 ml
CK Substrate Ac-LR-AFC (10 mM) 0.2 ml
CK Inhibitor (1 mM) 20 ul
Detection method :
Fluorescence (Ex/Em 400/505)
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