NucView™488 and RedDot™2 Apoptosis/Necrosis Assay Kit 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. RedDot™2 has far-red emission at 695 nm, well-separated from the green fluorescence of NucView™488. The excitation maximum of RedDot™2 is 665 nm, but it can be efficiently excited by wavelengths from 488 to 647 nm, and therefore can be used with the 488 nm flow cytometry laser line.
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