Caspase Activity Assays

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 offer a series of caspase assays that allow the simple detection of active caspases in living cells in real time or as a snapshot.

Select caspase activity assays

Also see flow cytometry assays for caspase activity
Also see microplate assays for caspase activity

Caspase Imaging Assays

  • Real-time no-wash assay for caspase activity
  • Multiparameter assays for live and dead cells
  • Antibodies for immunodetection of apoptosis

During apoptosis, ICE family members such as caspase-3 and -7 cleave PARP to yield an 85 kDa and a 25 kDa fragment. PARP cleavage is considered to be one of the classical characteristics of apoptosis. Antibodies to the 85 kDa fragment of cleaved PARP or to caspase-3 both serve as markers for apoptotic cells that can be monitored using immunofluorescence.

High-performance multiparameter apoptosis kits are formulated to allow the determination of live and dead cells in addition to the caspase status.

Select caspase imaging assays

Multiplex imaging of U2OS cells
Multiplex imaging of U2OS cells using CellEvent Caspase-3/7 Green detection reagent to detect apoptosis.

Caspase Assays in Microplates

  • Rhodamine 110-derived caspase substrates are available with a variety of enzyme specificities.
  • Real-time live-cell assays
  • Optimized kits for cell lysates
  • Individual substrates with choice of specificity

We offer microplate assays for caspase activity that can be performed on whole cells or on cell lysates. CellEvent Caspase-3/7 Green Detection Reagent is optimized for apoptosis analysis with a simple no-wash protocol that helps preserve delicate apoptotic cells. Live cells in microplate wells can be monitored in real time for apoptotic activity.

Microplate kits for caspase-3 measurement provide a choice of substrates with different wavelength emissions: Rhodamine 110 or AMCA can be used to report activity.

Dose response curve with CellEvent Caspase

Selection guide for imaging caspase activity

  CellEvent Caspase-3/7 Green Detection Reagent Image-iT LIVE Red Detection Kits, for microscopy Image-iT LIVE Green Detection Kits, for microscopy
Readout No-wash assay to monitor ongoing caspase activity. Fluorescence intensity increases with caspase activity. Assay kits based on signal retention require washing and provide a snapshot of caspase activity. Each kit format provides simultaneous detection of apoptotic cells (based on caspase activity), live cells, and dead cells.
Caspase Target Caspase-3/7 Poly-caspase Caspase-3/7 Poly-caspase Caspase-3/7
Fluorophore Reporter DEVD conjugated to a nucleic acid-binding dye SR-VAD-FMK SR-DEVD-FMK FAM-VAD-FMK FAM-DEVD-FMK
Standard filter sets FITC TRITC FITC
Em/Ex (nm) 502/530 550/595 488/530
Nuclear Counterstains included Not included Hoechst 33342 and SYTOX Green Hoechst 33342 and propidium iodide
Fluorescence Microscope Yes Yes Yes
High Content Instrument Yes Yes Yes
Microplate Reader Yes No No
Live Cells Yes Yes Yes
Fixed Cells No No No
Fixable Yes Yes Yes
Protocol outline
  1. Add diluted CellEvent Caspase-3/7 Green Detection Reagent to cells
  2. Incubate for 30 minutes 
  3. Measure fluorescence
  1. Add diluted reagent to cells
  2. Incubate for 60 minutes 
  3. Optional - stain with included nuclear counterstains 
  4. Wash
  5. Image
Bibliography Citations
Format 100 µl of 2 mM reagent in DMSO 25 µl of 2 mM reagent in DMSO ReadyProbes version in Dropper Bottles 25-assay kit
Cat. No. C10423 C10723 R37111 I35101 I35102 I35104 I35106