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. Life Technologies offers 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.

Multiparameter apoptosis kits are formulated to allow the determination of live and dead cells in addition to the caspase status.

  Detecting apoptosis with CellEvent™ Caspase-3/7 Green detection reagent
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

Life Technologies offers 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-3/7 Green Detection Reagent.
Dose response curve with CellEvent™ Caspase-3/7 Green Detection Reagent.


Selection Guides for Caspase Assays

 
Readout The easiest fluorescent staining method for apoptosis; no calculations, no dilutions, no pipetting
Caspase 3,7
Standard filter set FITC
Ex/Em (nm) 502/530
Bibliography Citations
Fluorescence microscope Yes
HCS Yes
HTS Yes
Live cells Yes
Fixed cells No
Fixable Yes
Format 6 x dropper bottles
Cat. No. R37111
  PARP FITC Apoptosis Kit CASP3 [D175] Monoclonal Antibody (clone 9H19L2), ABfinity™ Recombinant
Readout No-wash assay to monitor ongoing caspase activity. Fluorescence intensity increases with caspase activity. Marker for detecting apoptotic cells. Anti-Casp3 enables caspase detection in fixed and permeabilized cells
Caspase 3,7 3,7 3
Standard filter set FITC FITC Unconjugated
Ex/Em (nm) 502/530 490/525 Rabbit IgG
Bibliography Citations Citations Citations
Fluorescence microscope Yes Yes Yes
HCS Yes Yes Yes
HTS Yes    
Live cells Yes No No
Fixed cells No Yes Yes
Fixable Yes Yes Yes
Format 100 µL 1 kit, 100 tests 100 µg
Cat. No. C10423 AHM2011 700182
 
Readout 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.
Standard filter sets DAPI FITC TRITC DAPI FITC Texas Red®
Fluorophore Hoechst 33342 SYTOX® Green FMK Hoechst 33342 FAM Propidium iodide
Ex/Em (nm) 350/461 504/523 550/595 350/461 488/530 535/617
Bibliography Citations
Fluorescence microscope Yes Yes
HCS Yes Yes
HTS    
Live cells Yes Yes
Fixed cells No No
Fixable Yes Yes
Caspase Poly-caspase 3,7 Poly-caspase 8 3,7
Format 25-assay kit 25-assay kit 25-assay kit 25-assay kit 25-assay kit
Cat. No. I35101 I35102 I35104 I35105 I35106