Uses
Ac-DEVD-AFC is a fluorogenic substrate for caspase-3 (CPP32) and related cysteine proteases, which emits a blue to green shift in fluorescence. It also acts as a substrate for granzyme B-processed caspase-6, caspase-7, caspase-8, and caspase-10. It is known that Ac-DEVD-AFC has a greater Stokes'' shift than Ac-DEVD-AMC once it has been cleaved. Since caspase-3 is an effector caspase, it is involved in the cleavage of many substrates which promote apoptosis. This indicator excites light at 400 nm and emits light at 505 nm. In addition, Ac-DEVD-AFC is modeled after Asp216 on caspase-3, the PARP cleavage site. Ac-DEVD-AFC is an essential indicator used to monitor the cellular effects of caspase-3.
Biological Activity
ac-devd-afc is a fluorogenic substrate for activated caspase-3 (cpp32) with km value of 9.7 μm, and related cysteine proteases [1][2].apoptosis is a process of programmed cell death that occurs in multicellular organisms. caspase are a family of protease enzymes playing essential roles in programmed cell death (including apoptosis, pyroptosis and necroptosis) and inflammation. caspase activation is a major event in apoptosis. caspase-3 cleaves and activates caspases 6 and 7, and is processed and activated by caspases 8, 9, and 10 [1][2][3].ac-devd-afc (n-acetyl-asp-glu-val-asp-7-amido-4-trifluoromethylcoumarin) is a fluorogenic substrate for activated caspase-3 and related caspases. during apoptosis, activated caspase-3 cleaves poly (adp-ribose) polymerase, which it specifically targets at the amino sequence asp-glu-val-asp (devd). sequence is based on parp cleavage at asp216 for caspase-3 [1][2]. caspase activity can be quantified by fluorescent detection of free afc (7-amino-4-trifluoromethylcoumarin) at excitation 400 nm and emission 505 nm [2].
Biochem/physiol Actions
A fluorogenic substrate for caspase 3, a protease that is rapidly activated when cells are exposed to apoptotic conditions and that cleaves poly(ADP-ribose) polymerase. The 7-amido-4-trifluoromethylcoumarin derivative has better membrane permeability than the 7-amido-4-methylcoumarin derivative (A1086).
References
[1]. lazebnik ya1, kaufmann sh, desnoyers s, et al. cleavage of poly(adp-ribose) polymerase by a proteinase with properties like ice. nature. 1994 sep 22;371(6495):346-7.
[2]. xiang, j.,chao, d.t., and korsmeyer, s.j. bax-induced cell death may not require interleukin 1β-converting enzyme-like proteases. proceedings of the national academy of sciences of the united states of america 93, 14559-14563 (1996).
