| NucView 405 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 caspase-3 substrates can be used to detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. 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. The staining is also formaldehyde-fixable. 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 405 Caspase-3 Substrate also is offered 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 405 staining. Biotium also offers green fluorescent NucView 488 caspase-3 substrate and kits, as well as orange fluorescent NucView 530. Pacific Blue is a registered trademark of Thermo Fisher Scientific. |
