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-H Activity BioAssay(TM) 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.
Kit Components:
CH Cell Lysis Buffer, 1x25ml
CH Reaction Buffer, 1x5ml
CH Substrate R-AFC, 10mM, 1x0.2ml
CH 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.