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-D Activity Assay kit is a fluorescence-based assay that utilizes the preferred cathepsin-D substrate sequence GKPILFFRLK(Dnp)-D-R-NH2) labeled with MCA. Cell lysates or other samples that contain cathepsin-D will cleave the synthetic substrate to release fluorescence, which can then easily be quantified using a fluorometer or fluorescence plate reader at Ex/Em = 328/460 nm.
|Detection Method||Fluorescence (Ex/Em 328/460)|
|Applications||Detects alternative proteolytic enzymes such as the lysosomal cathepsin proteases that can initiate or propagate proapoptotic signals.|
|Features & Benefits||• Simple one-step procedure; takes only 1-2 hours
• Fast and convenient
• Cathepsin-D assay is straightforward, and can be adapted to 96-well plate assays.
|Kit Components||• CD Cell Lysis Buffer
• CD Reaction Buffer
• CD Substrate (1mM)
-20°C;shipping with Gel pack
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