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General description
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.
Application
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:
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
Quantity
The kit provides sufficient reagents for 100 tests of 100 μL in 96-well plates.
Kit Components Only
Product No.
Description
- N-Acetyl-Ile-Glu-Thr-Asp-al, ≥97% (HPLC) 100 μL
- Acetyl-Ile-Glu-Thr-Asp-p-nitroaniline Substrate 100 μL
- 10X Assay Buffer 5 mL
- 5X Lysis Buffer 5 mL
Storage Class Code
10 - Combustible liquids
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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FEBS letters, 583(4), 848-854 (2009-02-27)
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Molecular Cancer Therapeutics, 5(12), 3197-3197 (2006)
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Basic Concepts of Molecular Pathology, 32-32 (2009)
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