Caspase Assays for Flow Cytometry
A distinctive feature of the early stages of apoptosis is the activation of caspase enzymes, which participate in the cleavage of protein substrates and in the subsequent disassembly of the cell. We provide a series of caspase assays that allow the simple detection of active caspases in living cells by flow cytometry.
Real-time monitoring with the no-wash CellEvent Caspase-3/7 assay
The CellEvent Caspase-3/7 Flow Cytometry Assay enables the flow cytometric detection of activated caspase-3 and caspase-7 in apoptotic cells using novel, fluorogenic caspase-3/7 detection reagents (Figure 1).
- Caspase-3/7 specific—includes the recognition sequence for activated caspase-3 and caspase-7
- Easy identification—can be used to clearly identify live, dead, and apoptotic cell populations
- Quick analysis—washing and fixation is not required
- Compatibility—available for 488 nm excitation
CellEvent Caspase-3/7 detection reagent is cell-permeant. The reagent consists of a four-amino acid peptide (DEVD) conjugated to a nucleic acid-binding dye. During apoptosis, caspase-3 and caspase-7 proteins are activated and able to cleave the caspase 3/7 recognition sequence encoded in the DEVD peptide. Cleavage of the recognition sequence and binding of DNA by the reagent labels the apoptotic cells with a bright, fluorogenic signal that has absorption/emission maxima of ~511/533 nm for the green detection reagent. When used together with the appropriate SYTOX dead cell stain, apoptotic cells can be easily discriminated from live and necrotic cells.