Features of these indicators include:
• Optimized caspase-3/7 substrate for apoptosis analysis
• Easy to use—just add and read; no cell lysis required
>• For time course measurements; easy selection of target time points
• Compatible with live cell fluorescence imaging and formaldehyde fixation methods
• Supports multi-channel detection—combine with other fluorescent reagents to confirm the presence of Apoptosis

CellEvent™ Caspase-3/7 Green Detection Reagent:Maximum absorption/emission wavelength ∼502/530 nm. Detection can be performed using the FITC/GFP conventional filter set.

CellEvent™ Caspase-3/7 Red Detection Reagent: Maximum absorption/emission wavelength ∼590/610 nm. Detection can be performed using the traditional Texas red filter set.

CellEvent™ Caspase-3/7 Red Detection Reagent and CellEvent™ Caspase-3/7 Green Detection Reagent are supplied in dry powder form. Prepare 100X stock solution by simply adding 100µL PBS to the dry powder. Dilute this 100X stock 1:100 in complete medium and stain cells for 30-60 minutes before analyzing with a fluorescence microscope.

CellEvent™ Caspase-3/7 Red Detection Reagent is also available as a ready-to-use solution in DMSO. DMSO solution is a 400X ready-to-use stock solution. Simply dilute this stock 1:400 with growth medium and stain cells for 30-60 minutes.

To prevent apoptotic cells from being washed away, there is no need to wash at the end of staining. Stained cells can be subjected to live analysis or fixed with a 4% paraformaldehyde solution, such as Image-IT™ Paraformaldehyde 4% in 0.1M phosphate buffer (pH 7.4) (Cat. No. I28800). These fixed cells can be further processed for use in ICC workflows.

Substrate Specificity
CellEvent Caspase-3/7 Detection Reagent is a four-amino acid peptide (DEVD) conjugated to a nucleic acid-binding dye. The DEVD peptide sequence is the cleavage site for caspase-3/7, and the coupled dye does not fluoresce until it is cleaved from the peptide and bound to DNA.

Mechanism of Action
CellEvent Caspase-3/7 Green Detection Reagent itself does not fluoresce due to the DEVD peptide inhibiting the dye's ability to bind to DNA. However, upon activation of caspase-3/7 in apoptotic cells, the DEVD peptide is cleaved, allowing the dye to bind to DNA and react to produce bright fluorescence.

Simple three-step protocol
To use CellEvent Caspase 3/7 Assay Reagent, simply add it to cells and incubate for 30–60 minutes. Observations are available. Apoptotic cells containing activated caspase-3/7 have bright green or red nuclei, whereas cells without activated caspase-3/7 emit only a weak fluorescent signal.

Assay versatility enables detection and fixation of living cells
This robust assay enables researchers to detect caspase-3/ 7 activation is checked. Furthermore, since the washing step is not required for the assay, fragile apoptotic cells that are usually lost during the washing step are preserved. Importantly, the fluorescent signal from the CellEvent Caspase 3/7 Assay Reagent is preserved during fixation and permeabilization. This allows the flexibility to perform endpoint assays and probe other proteins of interest using immunocytochemistry.