NucView® 530 Caspase-3 Substrate is a convenient tool for detecting apoptosis in intact cells based on caspase-3/7 activity using confocal microscopy, flow cytometry, or live cell imaging. NucView® 530 is also available as 1 mM in PBS (10408) for use with DMSO-sensitive cell types.
- Orange fluorescent dye
- 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
To learn about the advantages of monitoring apoptosis using NucView® caspase-3 substrates, visit the NucView® Technology Page.
NucView® 530 Caspase-3 Substrate stains apoptotic cell nuclei with orange fluorescence, 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® 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 can be used to detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. Staining is compatible with subsequent fixation and permeabilization for immunostaining.
NucView® substrate are offered as solutions in DMSO or phosphate-buffered saline (PBS). The substrates in PBS are 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® staining.
NucView is a registered trademark of Biotium. NucView enzyme substrate technology is covered by U.S. patents. Cy dye is a registered trademark of GE Healthcare.
Find a list of NucView® references and a list of validated cell lines under Supporting Documents.