Product Pathways - Apoptosis / Autophagy
Caspase-7 Activity Assay Kit #9850
| Kit Includes | Quantity |
|---|---|
| Caspase-7 Antibody | 150 microliters |
| Chaps Cell Extract Buffer (10X) # 9852 | 1.5 milliliters |
| Fluorogenic Caspase-7 Substrate | 0.1 milliliters |
| 1M DTT | 1 milliliters |
| 10X Reaction Buffer | 15 milliliters |
Molecular Weight
Expected Molecular Weight: 35, 20 kDa
Reactivity
H
Reactivity Key: H=Human
Description
The Caspase-7 Activity Assay Kit includes all reagents necessary to assay caspase-7 activity, including Caspase-7 Antibody, fluorescent caspase substrate, reaction buffer and cell extract buffer. The assay system uses Caspase-7 Antibody to selectively immunoprecipitate caspase-7. Fluorogenic substrate is added and is cleaved proportionally to the amount of activated caspase-7, generating free fluorescent AFC. Free AFC is measured fluorometrically at lmax = 505 nM. Compared to other commercially available caspase-7 activity assays, this kit is designed to be more specific by using Caspase-7 Antibody to selectively immunoprecipitate caspase-7 from cell lysates.
Table
Titration curve of Caspase-7 activity assay using varying amounts of Jurkat cells, untreated or etoposide-treated (6 hours). Cell extracts were prepared for immunoprecipitation using Caspase-7 Antibody. Immune complexes were incubated with peptide substrate Ac-DEVD-AFC. Specific Caspase-7 activity was measured based on the fluorescent AFC group from the hydrolysis of Ac-DEVD-AFC by activated Caspase-7. Specific inhibition assay was performed by preincubating the immune complex with Ac-DEVD-CHO for 30 minutes before adding Ac-DEVD-AFC.
Background
Caspase-7 (CMH-1, Mch3, ICE-LAP3) has been identified as a major contributor to the execution of apoptosis (2-4). Caspase-7 is an effector caspase (along with caspase-2 and -3), meaning that it cleaves essential cellular machinery rather than activating other caspases (5-8). Caspase-7 is cleaved by many enzymes, including caspases-3, -6, -8, -9 and granzyme B (1,4,5). Once activated, caspase-7 cleaves many of the same substrates as caspase-3, including poly (ADP-ribose) polymerase, or PARP (2,4).
- Fernandes-Alnemri, T. et al. (1995) Cancer Res. 55, 6045-6052.
- Duan, H. et al. (1996) J. Biol. Chem. 271, 1621-1625.
- Lippke, J. A. et al. (1996) J. Biol. Chem. 271, 1825-1828.
- Cohen, G.M. (1997) Biochem. J. 326, 1-16.
- Thornberry, N. A. et al. (1997) J. Biol. Chem. 272, 17907-17911.
- Chandler, J. M. et al. (1998) J. Biol. Chem. 273, 10815-10818.
- MacFarlane, M. et al. (1997) J. Cell Biol. 137, 469-479.
- Nu?ez, G. et al. (1998) Oncogene 17, 3237-3245.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
