Ac-DEVD-AFC (Synonyms: N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-Trifluoromethylcoumarin,Caspase-3 Substrate (Fluorogenic)) |
| Catalog No.GC17602 |
fluorogenic substrate for activated caspase-3
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 201608-14-2
Sample solution is provided at 25 µL, 10mM.
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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).
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. | |
| Cas No. | 201608-14-2 | SDF | |
| 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 | ||
| Canonical 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 | ||
| Formula | C30H34F3N5O13 | M.Wt | 729.6 |
| Solubility | ≥ 73mg/mL in DMSO, <7.29mg/mL in Water | Storage | -20°C, protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.3706 mL | 6.8531 mL | 13.7061 mL |
| 5 mM | 0.2741 mL | 1.3706 mL | 2.7412 mL |
| 10 mM | 0.1371 mL | 0.6853 mL | 1.3706 mL |
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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
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- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
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Average Rating: 5 (Based on Reviews and 10 reference(s) in Google Scholar.)
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