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 fluorescence labeling^[2]
(1) Preparation of caspase buffer: 0.5% Igepal CA-630, 10 mM HEPES (pH 7.4), 2 mM EDTA, 0.5 mM phenylmethylsulphonyl fluoride (PMSF) and 5 μg/mL leupeptin.
(2) PC12 cells were treated with 50 μM 6-OHDA for 20 h.
(3) Collect cells by centrifugation at 250 g for 5 min, wash with PBS, and then centrifuge at 250 g for 5 min.
(4) Resuspension cells with caspase buffer at a ratio of 2 μL/10⁵ cells and lyse at 4°C for 20 min.
(5) Collect the lysate by centrifugation at 7200 g, 4°C and 10 min.
(6) Incubate 20 μL lysate with 100 μM Ac-DEVD-AMC at 30°C for 10 min.
(7) Measure caspase activity using a fluorescence spectrophotometer at an excitation wavelength of 380 nm and an emission wavelength of 460 nm.

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

675.64

C₃₀H₃₇N₅O₁₃

169332-61-0

Solid

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)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [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]