GLPBio

Ac-DEVD-AFC

Catalog No.: GC17602Purity:>99.00%Synonyms: N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-Trifluoromethylcoumarin,Caspase-3 Substrate (Fluorogenic)
COASDSData Sheet
fluorogenic substrate for activated caspase-3

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Ac-DEVD-AFC
Cas No.: 201608-14-2
1mgIn Stock
$54.00
1
5mgIn Stock
$162.00
1
10mgIn Stock
$297.00
1
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Description

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].

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).

Protocol

Cell experiment:

For the detection of caspase-3 activity, PBS washes cell pellets (derive from either the medium or the adherent cells) which are suspended in extract buffer [25 mM HEPES (pH7.4), 0.1% TritonX-l00, 10% glycerol, 5 mM DTT, 1mM phenylmethylsulfonyl fluoride, 10 mg/mL pepstatin, and 10 mg/mL Leupeptin] and vortexed vigorously. 20μl of extract (corresponding to 10% of the sample) are incubated with the caspase-3 fluorogenic substrates Ac-DEVD-AFC at 100 μM final concentration at room temperature, and caspase-3 activity is measured continuously by monitoring the release of fluorigenic AFC at 37°C[1].

References:

[1]. Wang X, et al. Involvement of Bim in Photofrin-mediated photodynamically induced apoptosis. Cell Physiol Biochem. 2015;35(4):1527-36.

Product Documents

Purity:>99.00%Appearance:A solid
COASDSData Sheet

Chemical Properties

CAS No.
201608-14-2
Synonyms
N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-Trifluoromethylcoumarin,Caspase-3 Substrate (Fluorogenic)
Chemical Name
N-acetyl-L-α-aspartyl-L-α-glutamyl-L-valyl-N-[2-oxo-4-(trifluoromethyl)-2H-1-benzopyran-7-yl]-L-α-asparagine
SMILES
CC(N[C@@H](CC(O)=O)C(N[C@H](C(N[C@@H](C(C)C)C(N[C@@H](CC(O)=O)C(NC1=CC(OC(C=C2C(F)(F)F)=O)=C2C=C1)=O)=O)=O)CCC(O)=O)=O)=O
Molecular Formula
C30H34F3N5O13
Molecular Weight
729.6
Solubility
≥ 73mg/mL in DMSO, <7.29mg/mL in Water
Storage
-20°C, protect from light
General tips
Select a suitable solvent according to the product solubility data. Once prepared, aliquot the stock solution to avoid repeated freeze-thaw cycles.
Use stock solutions within 6 months when stored at -80°C or within 1 month when stored at -20°C.
To improve solubility, warm the tube to 37°C and sonicate it for an appropriate period.
Shipping Condition
Evaluation samples are shipped with blue ice. Other available sizes are shipped at room temperature or with blue ice upon request.

CalculatorsMolarity / Dilution / Molecular Weight / Unit Conversion / In Vivo Formulation / Solubility

g/mol

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