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. 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.

High-performance 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

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


Selection Guides for Imaging Caspase Activity

 
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
 
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
  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