For detection of active caspase-3 in mammalian cells undergoing apoptosis
Caspase-3 is a proteolytic enzyme with a substrate selectivity for the peptide sequence Asp-Glu-Val-Asp (DEVD). FITC-C6-DEVD-FMK is a non-toxic, cell-permeable fluorescent substrate that irreversibly binds to activated caspase-3 in apoptotic cells. The FITC-labeled substrate can be detected with high sensitivity using flow cytometry.
Caspases are a family of proteases that play a central role in cellular apoptosis through the cleavage of select proteins, and result in the disassembly of the cell (Thornberry & Lazebnik Science, 1998). This process is switched on by pro-apoptotic signals from death receptors or stimuli such as cytotoxic reagents, which activate the initiator caspases and lead to activation of the effector caspases—caspase-3 and caspase-7 (Thornberry & Lazebnik Science, 1998). FITC-C6-DEVD-FMK is useful for screening caspase-3 inhibitors and quantifying apoptotic cells containing active caspase-3.
Figure 1. FITC-C6-DEVD-FMK
Jurkat cells were treated with 0.2% DMSO (Control) or 20 µM camptothecin (CPT), and in the presence or absence of Z-VAD-FMK (a pan-caspase inhibitor). Active caspase-3/7 was detected by flow cytometry, using FITC-C6-DEVD-FMK. A representative plot of three independent experiments is depicted in the figure above.
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