Caspase-8 Colorimetric Assay Kit
This Caspase-8 Colorimetric Assay Kit is designed to detect caspase-8 cleaving activity, Caspase-8 (FLICE/MACH-1) is a 55 kDa cytosolic protein with homology to the CD95/Fas-associated signal transduction molecule, FADD, in addition to its homology with other caspases. The kit contains a caspase-8 fluorogenic substrate (Ac-IETD-pNA) and assay buffers. The assay is based on spectrophotometric detection of the chromophore p-nitroanilide (pNA) after cleavage from the labeled substrate IETD-pNA. The pNA light emission can be quantified using a
spectrophotometer or a microtiter plate reader at 400- or 405-nm. Comparison of the absorbance of pNA from an apoptotic sample with an uninduced control allows determination of the fold increase in FLICE activity.
IETD-pNA Substrate: 500 μL
1 M dithiothreitol (DTT): 400 μL
Lysis Buffer: 100 mL
2X Reaction Buffer: 10 ml
Store at ≤ -20 °C for up to 6 months after initial use. Avoid repeated freeze-thaw cycles.
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