NucView(R) 530 Caspase-3 Substrate, 1 mM in 1X PBS

NucView(R) 530 Caspase-3 Substrate, 1 mM in PBS, provides a convenient tool for detecting apoptosis in intact cells based on caspase-3/7 activity using either confocal microscopy or flow cytometry. In contrast to other fluorogenic caspase substrates or fluorescent caspase inhibitor based (FLICA) assays, NucView(R) caspase-3 substrates can be used to detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. NucView(R) 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(R) Caspase-3 Substrates are bifunctional, allowing detection of caspase-3/7 activity and visualization of morphological changes in the nucleus during apoptosis. The staining is also formaldehyde-fixable.  NucView(R) 530 Caspase-3 Substrate stains apoptotic cell nuclei with orange fluorescence (Ex/Em 528/563 nm), for detection in the Cy(R)3 channel by fluorescence microscopy (Figure 2) or the PE channel by flow cytometry. NucView(R) 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(R) 530 also fluoresces in the FITC channel, and therefore cannot be analyzed together with green probes by flow cytometry.  NucView(R) 530 Caspase-3 Substrate also is available as a 1 mM solution in DMSO. 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(R) 530 staining. Biotium also offers blue fluorogenic NucView(R) 405 Caspase-3 Substrate, and green fluorogenic NucView(R) 488 Caspase-3 Substrate and kits.  Cy dye is a registered trademark of GE Healthcare.