Z-DEVD-FMK (Caspase-3 Inhibitor) is a selective, irreversible Caspase-3 inhibitor, and also exhibits effective inhibition activity on caspase-6, caspase-7, caspase-8, and caspase-10.

Caspase-3:18 μM.

METHODS: v-K-ras transformed normal rat kidney cells KNRK were treated with SCH56582 (20 µM) and Z-DEVD-FMK (20-50 µM) for 24 h. Cell death was detected using trypan blue.
RESULTS: Addition of 50 µM Z-DEVD-fmk resulted in >70% inhibition of apoptosis. [1]
METHODS: Human multiple myeloma cells KM3 were treated with betulinic acid (15 µg/mL) and Z-DEVD-FMK (50 mol/L) for 24 h, and the expression level of target proteins was detected using Western Blot.
RESULTS: Z-DEVD-FMK attenuated the betulinic acid-induced activation of caspase 3. Z-DEVD-FMK significantly blocked the cleavage of PARP. Z-DEVD-FMK significantly blocked PARP cleavage. [2]

METHODS: To test the in vivo activity, Z-DEVD-FMK (1.8 mg/kg in ethanol and freshly diluted in PBS containing 2% Tween-80) was injected intraperitoneally into C57BL/6 mice, followed by CPT-11 (350 mg/kg) 30 min later. Z-DEVD-FMK was continued to be administered once daily for three days.
RESULTS: Caspase-3 inhibitor Z-DEVD-FMK attenuated PT-11-induced GATA6-deficient mouse peritoneal macrophages. [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)