Contains NucView 488 Caspase-3 Substrate for detection of caspase-3/7 activity.
The far-red dead-cell stain RedDot 2 is included for staining necrotic and late apoptotic cells that have compromised plasma membrane integrity. This kit provides a convenient tool for profiling apoptotic and necrotic cell populations by fluorescence microscopy or flow cytometry.
In contrast to other fluorogenic caspase substrates or fluorescent caspase inhibitor based (FLICA) assays, NucView 488 Caspase-3 Substrate can be used to detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. The substrate consists 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 bright green fluorescence. Thus, NucView 488 Caspase-3 Substrate allows detection caspase-3/7 activity and visualization of morphological changes in the nucleus during apoptosis.
RedDot 2 is a cell membrane-impermeable, far-red dye with high selectivity for membrane compromised or dead cells. The far-red emission of RedDot 2 is well-separated from the green fluorescence of NucView 488. RedDot 2 can be excited by wavelengths from 488 to 647 nm, and therefore can be used with the 488 nm flow cytometry laser line.