ViaStain™ Live Caspase 3/7 Detection for 2D/3D Culture kit is designed to detect caspase 3/7 activity in live cells. The ability to perform kinetic apoptosis assays allows researchers to continuously measure the caspase 3/7 activity within the cell population. This no wash assay has been shown to effectively work in both 2D and 3D cultures.
The reagent (NucViewTM) consists of a nucleic acid-binding dye with a fluorescent probe that is attached to a four-amino acid peptide sequence DEVD (Asp-Glu-Val-Asp) forming a cell membrane-permeable DEVD-DNA complex. While the nucleic-acid dye is linked to the DEVD peptide sequence, the dye is unable to bind to DNA and remains non-fluorescent. During apoptosis, caspase 3/7 proteins cleave the DEVD-DNA dye complex and thereby release the high-affinity DNA dye, which translocates to the nucleus and binds to the DNA, producing a bright green fluorescent signal.
To enumerate the total number of nucleated cells and obtain percent of apoptotic cells, the cells can be counterstained with Hoechst 33342.