NucView® 405 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® 405 is also available at 1 mM in PBS (10407), for use in DMSO-sensitive cells.
- Blue 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® 405 dye is excited by the 405 nm laser line for detection by confocal microscopy in the DAPI channel, or by flow cytometry in the Pacific Blue® channel. NucView® 405 is ideal for caspase-3 detection in multi-color applications for researchers who wish to reserve the green fluorescence channel for other detection reagents.
Note: NucView® 405 signal may be difficult to observe using epifluorescence microscopy. Confocal microscopy using 405 nm laser excitation is recommended for imaging.
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® 405 staining.
NucView is a registered trademark of Biotium. NucView enzyme substrate technology is covered by U.S. patents. Pacific Blue is a registered trademark of Thermo Fisher Scientific.
Find a list of NucView® references and a list of validated cell lines under Supporting Documents.