NucView® Caspase-3 Substrates can be added to the cells at the start of the experiment or at the end. A major advantage of NucView® Caspase-3 Substrates compared to other apoptosis assays is that it can be used to monitor capase-3 activity in real time.
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
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 is also available as 1 mM in PBS (catalog number 10408) for use with DMSO-sensitive cell types.