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.
Sample Type:
Cell and tissue lysates
Intended Use:
Detects alternative proteolytic enzymes such as the lysosomal cathepsin proteases that can initiate or propagate proapoptotic signals.
Test Principle:
The Cathepsin-L Activity BioAssay(TM) kit is a fluorescence-based assay that utilizes the preferred cathepsin-L substrate sequence FR labeled with AFC (amino-4-trifluoromethyl coumarin). Cell lysates or other samples that contain cathepsin-L will cleave the synthetic substrate FR-AFC to release free AFC.
Kit Components:
CL Cell Lysis Buffer, 1x25ml
CL Reaction Buffer, 1x5ml
CL Substrate Ac-FR-AFC, 10mM, 1x0.2ml
CL Inhibitor, 1mM, 1x20ul
Storage and Stability:
Store powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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