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Caspase 3 Assay Kit (Fluorometric) (ab39383) provides a simple and convenient means for assaying the DEVD-dependent caspase activity. The assay is based on detection of cleavage of substrate DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). DEVD-AFC emits blue light (λ max = 400 nm); upon cleavage of the substrate by CPP32 or related caspases, free AFC emits a yellow-green fluorescence (Ex/Em = 400/505 nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-3/CPP32 activity.
Due to the nature of the substrate, this assay also detects caspase 7 activity.
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Activation of ICE-family proteases/caspases initiates apoptosis or other cellular processes in mammalian cells.
|2X Reaction Buffer||4 x 2ml|
|Cell Lysis Buffer||1 x 100ml|
|DEVD AFC||1 x 500µl|
|DTT||1 x 400µl|
Our Abpromise guarantee covers the use of ab39383 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent concentration.|
Caspase 3 activity in Jurkat lysates (6.6 x10e5 cells) following 20 hour exposure to 2 uM Camptothecin (ab120115) or 10 ng/mL anti-Fas Ab (MBL). Background signal subtracted, duplicates +/- SD.