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Description

Caspases are a family of protease enzymes that play essential roles in programmed cell death, termed apoptosis. Caspase-3 is a caspase protein that interacts with caspase-8 and caspase-9 and sequential activation of plays a central role in the execution-phase of cellular apoptosis.The BioTracker NucView 530 Red Caspase-3 Dye provides a convenient live cell tool for profiling apoptotic cell populations based on caspase-3/7 activity using either fluorescence microscopy or flow cytometry. In contrast to other fluorogenic caspase substrates or fluorescent caspase inhibitor based (FLICA) assays, the BioTracker NucView 530 Red Caspase-3 Dye can be used to detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. The substrate consists of a fluorogenic DNA dye coupled to the caspase-3/7 DEVD recognition sequence. The substrate, which is initially non-fluorescent, penetrates the plasma membrane and enters the cytoplasm. In apoptotic cells, caspase-3/7 cleaves the substrate, releasing the high-affinity DNA dye, which migrates to the cell nucleus and stains DNA with bright red fluorescence. Thus, NucView 530 Caspase-3 Substrate is bifunctional, allowing detection of caspase-3/7 activity and visualization of morphological changes in the nucleus during apoptosis. The dye is formaldehyde-fixable and compatible with subsequent immunostaining. The dye is offered in DMSO and PBS (phosphate-buffered saline) formulations. The substrate in PBS is formulated for use in cells that are sensitive to DMSO toxicity. NucView 530 Caspase-3 Substrate stains apoptotic cell nuclei with orange fluorescence, for detection in the Cy3 channel by fluorescence microscopy or the PE channel by flow cytometry. Note that when excited by the 488nm laser line, NucView 530 also fluoresces in the FITC channel, and therefore cannot be analyzed together with green probes by flow cytometry.