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Z-LETD-FMK

Z-LETD-FMK is a cell permeable, irreversible caspase protease 8 inhibitor that binds to the catalytic site of caspase- 8, which plays an important role in the induction of apoptosis. The binding of Z-LETD-FMK to caspase-8 inhibits the activity of this protease, enabling it to inhibit apoptotic events associated with caspase-8 activation. For inhibition of apoptosis, Z-LETD-FMK should be added at the same time that apoptosis is induced. 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.

Z-LETD-FMK stock solution
Add appropriate amount of DMSO into Z-LETD-FMK to make 2-5 mM Z-LETD-FMK stock solution.
Note     Store the unused Z-LETD-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-LETD-FMK along with the treatment of your choice.
    Note     We recommend using 10-100 μM of final concentration of Z-LETD-FMK to inhibit 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 your choice of instrumentation. 

References

View all 8 references: Citation Explorer
Rapamycin, an mTOR inhibitor, induced apoptosis via independent mitochondrial and death receptor pathway in retinoblastoma Y79 cell.
Authors: Wang, Yan-Dong and Su, Yong-Jing and Li, Jian-Ying and Yao, Xiang-Chao and Liang, Guang-Jiang
Journal: International journal of clinical and experimental medicine (2015): 10723-30
A polysaccharide from Sanguisorbae radix induces caspase-dependent apoptosis in human leukemia HL-60 cells.
Authors: Wu, Zhigang and Sun, Honghui and Li, Jingzhong and Ma, Chijiao and Zhao, Siqiao and Guo, Zheng and Lin, Yao and Lin, Yaping and Liu, Li
Journal: International journal of biological macromolecules (2014): 615-20
Use of fluorochrome-labeled inhibitors of caspases to detect neuronal apoptosis in the whole-mounted lamprey brain after spinal cord injury.
Authors: Barreiro-Iglesias, Antón and Shifman, Michael I
Journal: Enzyme research (2012): 835731
Caspase-8 and p38MAPK in DATS-induced apoptosis of human CNE2 cells.
Authors: Ji, C and Ren, F and Xu, M
Journal: Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas (2010): 821-7
[Synergistic effects and mechanisms of combined tumor necrosis factor-related apoptosis-inducing ligand and chemotherapeutic drugs or radiotherapy in killing laryngeal squamous carcinoma cells in vitro].
Authors: Zhang, Ming and Zhou, Liang
Journal: Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery (2009): 565-70
Page updated on December 17, 2024

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Catalog Number13307
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Fax1-800-609-2943
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Physical properties

Molecular weight

654.69

Solvent

DMSO

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12171501
Inhibition of caspase 8 Activities in Jurkat cells with caspase 8 inhibitor Z-LETD-FMK (Cat.# 13307). Jurkat cells were seeded on the same day at 200,000 cells/90 &micro;L/well in a Costar black wall/clear bottom 96-well plate. The cells were treated with or without 1 &micro;M of staurosporine for 5 hours. The Z-LETD-FMK was added and co-incubated with staurosporine. The caspase-8 activity was detected with Cell Meter&trade; 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 8 Activities in Jurkat cells with caspase 8 inhibitor Z-LETD-FMK (Cat.# 13307). Jurkat cells were seeded on the same day at 200,000 cells/90 &micro;L/well in a Costar black wall/clear bottom 96-well plate. The cells were treated with or without 1 &micro;M of staurosporine for 5 hours. The Z-LETD-FMK was added and co-incubated with staurosporine. The caspase-8 activity was detected with Cell Meter&trade; 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 8 Activities in Jurkat cells with caspase 8 inhibitor Z-LETD-FMK (Cat.# 13307). Jurkat cells were seeded on the same day at 200,000 cells/90 &micro;L/well in a Costar black wall/clear bottom 96-well plate. The cells were treated with or without 1 &micro;M of staurosporine for 5 hours. The Z-LETD-FMK was added and co-incubated with staurosporine. The caspase-8 activity was detected with Cell Meter&trade; 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).
Gallery Image 2

Documents

pdfSafety data sheet
pdfProduct protocol
pdfCertificate of analysis
Caspases