Caspase 8 Assay Kit, Colorimetric

Caspases (cysteine-requiring aspartate proteases) belong to a highly conserved family of cysteine proteases with specificity for aspartic acid residues found in their substrates. Caspases play a central role in apoptosis. Inhibition of caspases prevents the cell death process. Currently, there are 13 caspases recognized in mammals. Caspases are classified as initiators and effectors. Caspase 8 belongs to the class of initiators. It possesses a prodomain at the terminal and contains the death effector domain which serves as protein-protein interaction motif for caspase 8. With respect to substrate specificity, caspase 8 is classified as a member of group III. It specifically recognizes the (I/L/V) EXD sequence on the substrate.

The Caspase 8 Colorimetric Assay Kit is based on the hydrolysis of the peptide substrate Acetyl-Ile-Glu-Thr-Asp-p-Nitroaniline (Ac-IETD-pNA) by caspase-8 that results in the release of a p-Nitroaniline (p-NA) moiety. The p-NA is read at 405 nm. The concentration of the p-NA released can be calculated from a calibration curve prepared from p-NA standards (included with the kit).
The kit detects caspase 8 activity in crude and purified preparations of caspase 8.
Complete kit contains:

• purified caspase 8 for a positive control
• caspase 8 inhibitor
• p-NA fluorescent standard

The kit provides sufficient reagents for 100 tests of 100 μL in 96-well plates.