Ac-DEVD-AMC 

Ac-DEVD-AMC is a fluorescent substrate of caspase-3/caspase-7. When treating Ac-DEVD-AMC with cell lysate, Ac-DEVD-AMC releases amino-4-methylcoumarin (AMC) for fluorescence detection, with an excitation wavelength of 380 nm and an emission wavelength of 460 nm^[1][2][3].

Ac-DEVD-AMC荧光标记^[2]
(1) caspase缓冲液的制备：0.5% Igepal CA-630，10 mM HEPES (pH 7.4)，2 mM EDTA，0.5 mM苯甲基磺酰氟 (PMSF) 和 5 μg/mL 亮肽素。
(2) PC12 细胞用 50 μM 6-OHDA 处理 20 小时。
(3) 250 g 离心 5 分钟收集细胞，PBS洗涤，250g离心5分钟。
(4) caspase缓冲液按2uL/10⁵细胞重悬细胞，4℃裂解20 分钟。
(5) 通过在 7200 g、4°C 和 10 分钟下离心收集裂解物。
(6) 将 20 μL 裂解物与 100 μM Ac-DEVD-AMC 在 30°C 下孵育10 分钟。
(7) 使用荧光分光光度计在激发波长 380 nm 和发射波长 460 nm 下测量 caspase 活性。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

675.64

C₃₀H₃₇N₅O₁₃

169332-61-0

固体

White to off-white

480

390

N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-Methylcoumarin

Ac-DEVD-7-amido-4-Methylcoumarin

Room temperature in continental US; may vary elsewhere.

Sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

• [1]. Nobel CS, et al. Disulfiram is a potent inhibitor of proteases of the caspase family. Chem Res Toxicol. 1997 Dec;10(12):1319-24.  [Content Brief]

  [2]. Ochu EE, et al. Caspases mediate 6-hydroxydopamine-induced apoptosis but not necrosis in PC12 cells. J Neurochem. 1998 Jun;70(6):2637-40.  [Content Brief]

  [3]. Yakovlev AA, Gorokhovatsky AY, Onufriev MV, Beletsky IP, Gulyaeva NV. Brain cathepsin B cleaves a caspase substrate. Biochemistry (Mosc). 2008 Mar;73(3):332-6.  [Content Brief]