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.
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.
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.
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.|
|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|
|Nuclear Counterstains included||Not included||Hoechst 33342 and SYTOX Green||Hoechst 33342 and propidium iodide|
|High Content Instrument||Yes||Yes||Yes|
|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|
|Molecular Probes Handbook