21 Jul [ICT] Dual Sensor Casp 1 & MMP by Cell Technology
The Dual Sensor MitoCaspTM Detection Kit utilizes a cationic dye to visualize mitochondrial membrane potential and caspase activity simultaneously. The kit yields quantitative and qualitative results and can be used in conjunction with other antibodies or stains. Reagents are cell permeable.
Caspase enzymes specifically recognize a 4 amino acid sequence (on their substrate) which necessarily includes an aspartic acid residue. This residue is the target for the cleavage reaction, which occurs at the carbonyl end of the aspartic acid residue.
Caspases can be detected via immunoprecipitation, immuno-blotting techniques using caspase specific antibodies, or by employing fluorogenic substrates which become fluorescent upon cleavage by the caspase. MitoCasp uses a novel approach to detect active caspases.
The methodology is based on carboxyfluorescein (FAM) labeled fluoromethyl ketone (FMK)-peptide inhibitors of caspases. These inhibitors are cell permeable and non-cytotoxic. Once inside the cell, the inhibitor binds covalently to the active caspase. Cells that contain bound inhibitor can be analyzed by flow cytometry or fluorescence microscopy.
