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ApoSight™ Green Caspase 3/7 substrate
ApoSight™ Green Caspase 3/7 substrate is the first fluorogenic probe for the direct fluorescence imaing of caspase activities in live cells. It consists of three moieties including a). a masked fluorophore, b). a caspase-selective peptide fragment (DEVD), and c). a cell-penetrating moiety. The cell-penetrating moiety carries the probe into live cells. Upon entering live cells, the caspase-selective peptide fragment is cleaved by a caspase to release the masked fluorophore. The intensity of recovered fluorescence is directly related to the activity of the caspase 3/7 to be measured. Compared to the existing caspase assays in live cells, ApoSight™ Green Caspase 3/7 substrate is much more robust, convenient, and accurate. ApoSight™ Green Caspase 3/7 substrate releases a fluorophore that has Ex/Em ~490/520 nm. It does not need a DNA interaction to be fluorescent as reported for NucView reagents. It does not inhibit caspase activity as reported for the FMK peptide probes. Although fluorescent FMK peptide inhibitors of caspases are widely used for detecting caspase activities in live cells, this technology has a few severe limitations: a). FMK caspase inhibitors have high cytotoxicity since FMK peptides bind covalently to active caspases; b). The irreversible covalent binding of FMK peptides to caspases inhibits caspase activities, causing false positive apoptosis; c). FMK assays have extremely high background, and require intensive washings, resulting in extremely low throughput; d). FMK peptides are not stable in aqueous solutions and must be used immediately. ApoSight™ Green Caspase 3/7 substrate can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation.
The detection of caspase 3/7 activity in Jurkat cells with ApoSight™ Green Caspase 3/7 substrate. Jurkat cells (200,000 cells/well/ 96-well plate) were treated with 1 μM Staurosporine or DMSO for 4 hours. Cells were incubated with Caspase 3/7 Substrate working solution at 37°C for 1 hour. Images were taken with a fluorescence microscope using a FITC filter set.
The detection of caspase 3/7 activity in Jurkat cells with ApoSight™ Green Caspase 3/7 substrate. Jurkat cells (200,000 cells/well/ 96-well plate) were treated with 1 μM Staurosporine or DMSO for 4 hours. Cells were incubated with Caspase 3/7 Substrate working solution at 37°C for 1 hour. Images were taken with a fluorescence microscope using a FITC filter set.
protocol
Protocol
sds
SDS
COA
CatalogSize
Price
Quantity
13290100 Tests
Price
 
Physical properties
Molecular weight1290.61
SolventDMSO
Spectral properties
Extinction coefficient (cm -1 M -1)
80000
Excitation (nm)500
Emission (nm)522
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
Instrument settings
Flow cytometer
Excitation488 nm laser
Emission530/30 nm filter
Instrument specification(s)FITC channel
Fluorescence microscope
ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall/clear bottom
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Page updated on September 30, 2025