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Z-VAD-FMK

Documents
pdfSafety data sheet
pdfProduct protocol
pdfCertificate of analysis
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Show More (14)
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?
Application notes
Abbreviation of Common Chemical Compounds Related to Peptides
Annexin V
Calcein
Experimental Protocol for Calcein AM Assay
Human High Temperature Requirement Serine Protease A1 (HTRA1) Degrades Tau Protein Aggregates
AssayWise
Intracellular pH Measurement with Dual Excitation Fluorescence Sensor BCFL
Nucleic Acid Detection, Quantification and Imaging
A practical guide for use of PE and APC in flow cytometry
Calbryte™ series now available
Fluorescent Phalloidin: A Practical Stain for Visualizing Actin Filaments
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.
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).
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).
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).
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Catalog Number13300
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Request quotation
Physical properties
Molecular weight467.49
SolventDMSO
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
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.

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.
  1. Treat your samples as desired to induce apoptosis.
  2. 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.
  3. Stain samples as you desired.
  4. Monitor the fluorescence intensity with a proper instrument. 
Product family
NameExcitation (nm)Emission (nm)
FAM-VAD-FMK493517
TF4-VAD-FMK578602
SRB-VAD-FMK [Sulforhodamine B-VAD-FMK]559577
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
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
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
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
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
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