NucView® 530 Caspase-3 Substrate detects apoptosis in intact cells based on caspase-3/7 activity using confocal microscopy, flow cytometry, or live cell imaging.

• NucView® 530 dye has excitation/emission maxima of 528/563 nm (with DNA)
• Rapid, no-wash, endpoint or real-time assays
• Non-toxic, allowing multi-day experiments to be performed
• For flow cytometry, microscopy or live cell imaging systems
• Dual detection of caspase activity and nuclear morphology
• Formaldehyde fixable

NucView® substrates consist 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 fluorescence. Thus, NucView caspase-3 substrates are bifunctional, allowing detection of caspase-3/7 activity and visualization of morphological changes in the nucleus during apoptosis. In contrast to other fluorogenic caspase substrates or fluorescent caspase inhibitor based (FLICA) assays, NucView® caspase-3 substrates detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. Staining is compatible with subsequent fixation and permeabilization for immunostaining.

NucView® 530 Caspase-3 Substrate stains apoptotic cell nuclei with orange fluorescence (Ex/Em 528/563 nm), for detection in the Cy®3 channel by fluorescence microscopy or the PE channel by flow cytometry. NucView® 530 can be used for multi-color imaging with blue, green, or far-red fluorescent probes. Note that when excited by the 488 nm laser line, NucView® 530 also fluoresces in the FITC channel, and therefore cannot be analyzed together with green probes by flow cytometry.

NucView® 530 Caspase-3 Substrate is available as a 1 mM solution in DMSO (catalog no. 10406) or PBS (phosphate-buffered saline). The substrate in PBS is formulated for use in cells that are sensitive to DMSO toxicity. In non-DMSO sensitive cell types, adding DMSO during the substrate incubation may enhance NucView® 530 staining.

Download a full list of references: Validated cell lines for NucView® 488 with references

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