- Optimized caspase-3/7 substrate for apoptosis analysis
- Simple, no-wash protocol helps preserve delicate apoptotic cells
- Compatible with both live cell fluorescence-imaging and formaldehyde-based fixation methods
The CellEvent® Caspase-3/7 Green Detection Reagent is a four amino acid peptide (DEVD), conjugated to a nucleic acid binding dye with an absorption/emission maxima of ~502/530 nm. The DEVD peptide sequence is a cleavage site for caspase-3/7 and the conjugated dye is non-fluorescent until cleaved from the peptide and bound to DNA.
Principles of CellEvent® Caspase-3/7 Apoptosis Detection Reagent
The CellEvent® Caspase-3/7 Green Detection Reagent is intrinsically non-fluorescent as the DEVD peptide inhibits the ability of the dye to bind to DNA. However, after activation of caspase-3/7 in apoptotic cells, the DEVD peptide is cleaved enabling the dye to bind to DNA and produce a bright, fluorogenic response. The fluorescence emission of the dye when bound to DNA is ~530 nm and can be observed using a standard “FITC" filter set.
Simple Three Step Protocol
To use CellEvent® Caspase 3/7 Detection Reagent simply add substrate to cells, incubate 30 minutes, and visualize (Figure 1). Apoptotic cells with activated caspase-3/7 will have bright green nuclei, while cells without activated caspase 3/7 will have minimal fluorescence signal (Figure 2).
Assay Versatility Allows for Live Cell Detection and Fixation
Our robust assay enables researchers to examine caspase-3/7 activation in live cells. Furthermore, since wash steps are not necessary for detection, fragile apoptotic cells typically lost during wash steps are preserved. Importantly, the fluorescent signal from CellEvent® Caspase 3/7 Detection Reagent can survive fixation and permeabilization. This allows for the flexibility to perform end-point assays, and probe for other proteins of interest using immunocytochemistry.