CaspACE™ FITC-VAD-FMK In Situ Marker is a fluorescent analog of the pan caspase inhibitor Z-VAD-FMK (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone). The fluorescein isothiocyanate (FITC) group has been substituted for the carbobenzoxy (Z) N-terminal blocking group to create the fluorescent apoptosis marker. This structure allows delivery of the inhibitor into the cell where it irreversibly binds to activated caspases. The FITC label allows for a single-reagent addition to assay for caspase activity in situ. The FITC-VAD-FMK is supplied as a 5mM solution in DMSO and is intended for in situ monitoring of caspase activity by fluorescence detection. The suggested concentration for use in anti-Fas-treated Jurkat cell culture is 10μM.
Features and Benefits
A fast and simple protocol; just add FITC-VAD-FMK, incubate, wash and view fluorescence. No preparation of cell extracts or long incubation steps are necessary. The marker can be used with a variety of detection methods including fluorescence microscopy and flow cytometry and is adaptable to high-throughput applications. The CaspACE™ FITC-VAD-FMK In Situ Marker easily moves in and out of live cells and remains anchored inside cultured apoptotic cells.