Ac-DEVD-AFC
(Synonyms: AC-ASP-GLU-VAL-ASP-7-氨基-4-三氟甲基香豆素,N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-Trifluoromethylcoumarin,Caspase-3 Substrate (Fluorogenic)) 目录号 : GC17602
A fluorogenic caspase subsrate
Cas No.:201608-14-2
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
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. | |
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).
| Cas No. | 201608-14-2 | SDF | |
| 别名 | AC-ASP-GLU-VAL-ASP-7-氨基-4-三氟甲基香豆素,N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-Trifluoromethylcoumarin,Caspase-3 Substrate (Fluorogenic) | ||
| 化学名 | 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 | ||
| 分子式 | C30H34F3N5O13 | 分子量 | 729.6 |
| 溶解度 | ≥ 73mg/mL in DMSO, <7.29mg/mL in Water | 储存条件 | -20°C, protect from light |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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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 |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。