Z-DEVD-FMK (Caspase-3 Inhibitor) is a Caspase-3 inhibitor (IC50=18 μM) that is selective, irreversible, and cell-permeable. Z-DEVD-FMK inhibits apoptosis induced by caspase activation.

Caspase-3:18 μM.

方法：v-K-ras转化的正常大鼠肾脏细胞 KNRK 用 SCH56582 (20  µM) 和 Z-DEVD-FMK (20-50  µM) 处理 24 h，使用 trypan blue 检测细胞死亡。
结果：添加 50 µM Z-DEVD-fmk 导致细胞凋亡抑制 >70%。[1]
方法：人多发性骨髓瘤细胞 KM3 用 betulinic acid (15  µg/mL) 和 Z-DEVD-FMK (50  mol/L) 处理 24 h，使用 Western Blot 检测靶点蛋白表达水平。
结果：Z-DEVD-VMK 减轻了 betulinic acid 诱导的 caspase 3 的激活。Z-DEVD-FMK 显著阻断 PARP 的切割。[2]

方法：为测试体内活性，将 Z-DEVD-FMK (1.8 mg/kg in ethanol and freshly diluted in PBS containing 2% Tween-80) 腹腔注射给 C57BL/6 小鼠，30 min 后注射 CPT-11 (350 mg/kg)。Z-DEVD-FMK 继续给药三天，每天一次。
结果：Caspase-3 抑制剂 Z-DEVD-FMK 减弱 PT-11 诱导的 GATA6 缺失小鼠腹腔巨噬细胞。[3]

Caspase activity assay : Caspase-3 and caspase-9 activities are measured using fluorescent-based substrate. After treatment, the cells are resuspended in lysis buffer (50 mM Tris HCl, 1 mM EDTA, and 10 mM EGTA) containing 10 mM digitonin for 20 min at 37°C. Supernatants are treated with either of the fluorogenic substrates Ac-DEVD-AFC for caspase-3 or Ac-LEHD-AFC for caspase-9 for 1 h at 37°C and fluorescence is measured at excitation at 400 nm and emission at 505 nm using a Gemini XS fluorescence plate reade

N27 cells are incubated with 100 μM 6-OHDA for 24 h or 300 μM MPP+ for 36 h in the presence or absence of 50 μM Z-DEVD-FMK and cell death is determined by MTT (3-(4,5-dimethylthiazol-3-yl)-2,5-diphenyl tetrazolium bromide) assay, which is widely used to assess cell viability. After treatment, the cells are incubated in serum-free medium containing 0.25 mg/ml MTT for 3 h at 37°C. Formation of formazan from tetrazolium is measured at 570 nm with a reference wavelength at 630 nm using a SpectraMax microplate reader.(Only for Reference)