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Caspase 3 Assay Kit (Colorimetric) (ab39401) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence DEVD. The assay is based on spectrophotometric detection of the chromophore p-nitroaniline (p-NA) after cleavage from the labeled substrate DEVD-pNA. The p-NA light emission can be quantified using a spectrophotometer or a microtiter plate reader at 400- or 405 nm. Comparison of the absorbance of p-NA from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase 3 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 in mammalian cells.
|2X Reaction Buffer||4 x 2ml|
|Cell Lysis Buffer||1 x 100ml|
|DEVD-pNA||1 x 500µl|
|Dilution Buffer||1 x 100ml|
|DTT||1 x 400µl|
Image obtained from Xu G et al., JBC, 2012 Feb 11;28(7):4808-17
Caspase 3 in Jurkat lysates (3.3 x10e6 cells) following 20 hour exposure to 2 uM Camptothecin (ab120115) or 10 ng/mL anti-Fas Ab (MBL).
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