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Caspase 8 Assay Kit (Colorimetric) (ab39700) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence IETD. The assay is based on spectrophotometric detection of the chromophore p-nitroanilide (p-NA) after cleavage from the labeled substrate IETD-p-NA. 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 a treated sample with an untreated control allows determination of the fold increase in Caspase 3 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|
|Dilution Buffer||1 x 100ml|
|DTT||1 x 400µl|
|IETD-pNA||1 x 500µl|
Our Abpromise guarantee covers the use of ab39700 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.|
Active caspase 8 in control (CTRL) Jurkat cells (10e6/mL) or in cells after four hours exposure to 50 ng/mL anti-Fas Ab (αFas) (MBL), or pretreated one hour with 50 μM Z-VAD(OMe)-FMK (ab120487) followed by four hours with αFas. Background signal subtracted, duplicates; +/- SD.
ab39700 has not yet been referenced specifically in any publications.