Z-VAD-FMK
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | 467.49 |
Solvent | DMSO |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12171501 |
Overview | ![]() ![]() |
See also: Caspases
Molecular weight 467.49 |
Z-VAD-FMK (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone) is a cell-permeable, irreversible pan caspase inhibitor that binds to the catalytic site of caspase proteases and can inhibit induction of apoptosis. For inhibition of apoptosis, Z-VAD-FMK should be added at the same time that apoptosis is induced. The peptide is O-methylated in the P1 position on aspartic acid, providing enhanced stability and increased cell permeability. Fluoromethyl ketone (FMK)-derivatized peptides act as effective irreversible inhibitors with no added cytotoxic effects.
Example protocol
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
Note Store the unused Z-VAD-FMK stock solution at -20 °C in single use aliquots.
Z-VAD-FMK stock solution
Add appropriate amount of DMSO into Z-VAD-FMK to make 2-5 mM Z-VAD-FMK stock solution. Note Store the unused Z-VAD-FMK stock solution at -20 °C in single use aliquots.
SAMPLE EXPERIMENTAL PROTOCOL
The following protocol can be used as a guideline and should be optimized according to your needs.
- Treat your samples as desired to induce apoptosis.
- Add Z-VAD-FMK along with the treatment of your choice.
Note We recommend using 10-100 µM of final concentration ofZ-VAD-FMK to inhibit caspases such as 50 µM for caspase-8.
Note Optimal time and concentration for incubation needs to be determined experimentally. - Stain samples as you desired.
- Monitor the fluorescence intensity with a proper instrument.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of Z-VAD-FMK to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 213.908 µL | 1.07 mL | 2.139 mL | 10.695 mL | 21.391 mL |
5 mM | 42.782 µL | 213.908 µL | 427.817 µL | 2.139 mL | 4.278 mL |
10 mM | 21.391 µL | 106.954 µL | 213.908 µL | 1.07 mL | 2.139 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Product Family
Name | Excitation (nm) | Emission (nm) |
FAM-VAD-FMK | 493 | 517 |
TF4-VAD-FMK | 578 | 602 |
SRB-VAD-FMK [Sulforhodamine B-VAD-FMK] | 559 | 577 |
Images

Figure 1. Inhibition of caspase activities in Jurkat cells with pan caspase inhibitor Z-VAD-FMK (Cat.# 13300). Jurkat cells were seeded on the same day at 200,000 cells/90 µL/well in a Costar black wall/clear bottom 96-well plate. The cells were treated with or without 1 µM of staurosporine for 5 hours. The Z-VAD-FMK (50 uM) was added and co-incubated with staurosporine. The caspase-8 activity was detected with Cell Meter™ Caspase 8 Activity Apoptosis Assay Kit (Cat.# 22816). The fluorescence intensity was measured at Ex/Em = 540/620 nm (Cutoff = 610 nm) with Flexstation 3 fluorescence microplate reader (Molecular Devices).
References
View all 8 references: Citation Explorer
Identification of active caspases using affinity-based probes.
Authors: McStay, Gavin P and Green, Douglas R
Journal: Cold Spring Harbor protocols (2014): 856-60
Authors: McStay, Gavin P and Green, Douglas R
Journal: Cold Spring Harbor protocols (2014): 856-60
Induction of apoptosis in human renal cell carcinoma cells by vitamin E succinate in caspase-independent manner.
Authors: Wu, Xiu-Xian and Kakehi, Yoshiyuki and Jin, Xing-Hua and Inui, Masashi and Sugimoto, Mikio
Journal: Urology (2009): 193-9
Authors: Wu, Xiu-Xian and Kakehi, Yoshiyuki and Jin, Xing-Hua and Inui, Masashi and Sugimoto, Mikio
Journal: Urology (2009): 193-9
Metformin induces apoptosis of pancreatic cancer cells.
Authors: Wang, Luo-Wei and Li, Zhao-Shen and Zou, Duo-Wu and Jin, Zhen-Dong and Gao, Jun and Xu, Guo-Ming
Journal: World journal of gastroenterology (2008): 7192-8
Authors: Wang, Luo-Wei and Li, Zhao-Shen and Zou, Duo-Wu and Jin, Zhen-Dong and Gao, Jun and Xu, Guo-Ming
Journal: World journal of gastroenterology (2008): 7192-8
Induction of apoptosis in renal tubular cells by histone deacetylase inhibitors, a family of anticancer agents.
Authors: Dong, Guie and Wang, Lysa and Wang, Cong-Yi and Yang, Tianxin and Kumar, M Vijay and Dong, Zheng
Journal: The Journal of pharmacology and experimental therapeutics (2008): 978-84
Authors: Dong, Guie and Wang, Lysa and Wang, Cong-Yi and Yang, Tianxin and Kumar, M Vijay and Dong, Zheng
Journal: The Journal of pharmacology and experimental therapeutics (2008): 978-84
Apoptosis of gastric cancer cell line MKN45 by photodynamic treatment with photofrin.
Authors: Takahira, Kenichiro and Sano, Munetaka and Arai, Hajime and Hanai, Hiroyuki
Journal: Lasers in medical science (2004): 89-94
Authors: Takahira, Kenichiro and Sano, Munetaka and Arai, Hajime and Hanai, Hiroyuki
Journal: Lasers in medical science (2004): 89-94
Non-caspase-mediated apoptosis contributes to the potent cytotoxicity of the enediyne antibiotic lidamycin toward human tumor cells.
Authors: Wang, Zhen and He, Qiyang and Liang, Yunyan and Wang, Daishu and Li, Yi Yang and Li, Diandong
Journal: Biochemical pharmacology (2003): 1767-75
Authors: Wang, Zhen and He, Qiyang and Liang, Yunyan and Wang, Daishu and Li, Yi Yang and Li, Diandong
Journal: Biochemical pharmacology (2003): 1767-75
Androgen induced cell death in SHSY5Y neuroblastoma cells expressing wild-type and spinal bulbar muscular atrophy mutant androgen receptors.
Authors: Grierson, A J and Shaw, C E and Miller, C C
Journal: Biochimica et biophysica acta (2001): 13-20
Authors: Grierson, A J and Shaw, C E and Miller, C C
Journal: Biochimica et biophysica acta (2001): 13-20
Critical role for caspases 3 and 8 in neutrophil but not eosinophil apoptosis.
Authors: Daigle, I and Simon, H U
Journal: International archives of allergy and immunology (2001): 147-56
Authors: Daigle, I and Simon, H U
Journal: International archives of allergy and immunology (2001): 147-56
Application notes
FAQ
Are inflammasomes and caspase-1 related?
Do you offer any fluorimetric assays that measure caspase activation/activity in live cells using a flow cytometer?
Does pH and staining temperature affect Annexin V-Phosphatidylserine binding?
Does propidium iodide stain apoptotic cells?
How can I tell if my cell sample is dying?
Do you offer any fluorimetric assays that measure caspase activation/activity in live cells using a flow cytometer?
Does pH and staining temperature affect Annexin V-Phosphatidylserine binding?
Does propidium iodide stain apoptotic cells?
How can I tell if my cell sample is dying?