TF3-DEVD-FMK
| Overview |  SDSProtocol |
Molecular weight 975.04 | Correction Factor (280 nm) 0.179 | Extinction coefficient (cm -1 M -1) 750001 | Excitation (nm) 554 | Emission (nm) 578 |
TF3-DEVD-FMK is a superior replacement for 5-TAMRA-DEVD-FMK, i.e., 5-carboxytetramethylrhdamine-Asp-Glu-Val-Asp-fluoromethylketone since it can be better excited at 555 nm. TF3 dyes has identical spectral properties to Cy3 with significantly enhanced photostability. TF3-DEVD-FMK binds irreversibly to active caspase 3/7 in stimulated cells. The fluorescent intensity of the TF3-DEVD-FMK signal is proportional to the amount of active caspase 3/7 and can be easily detected by fluorescence microscopy, flow cytometer, or fluorescent microplate reader.
Example protocol
AT A GLANCE
Important notes
It is important to store at <-15 °C and should be stored in cool, dark place.
It can be used within 12 months from the date of receipt.
SAMPLE EXPERIMENTAL PROTOCOL
Following protocol only provides a guideline, and should be modified according to your specific needs.
General Solution Caspase Assays Using AMC, AFC, pNA, R110 and ProRed Substrates
- Prepare a 10 mM stock solution in DMSO.
- Prepare a 2X caspase substrate (50 µM) assay solution as the following: 50 µL substrate stock solution, 100 µL DTT (1M), 400 µL EDTA (100 mM), 10 mL Tris Buffer (20 mM), pH =7.4.
- Mix equal volume of the caspase standards or samples with 2X caspase substrate assay solution, and incubate the solutions at room temperature for at least 1 hour.
- Monitor the fluorescence using a fluorescence microplate reader, or absorbance using an absorbance microplate reader.
Cell Caspase Assays Using Cell-Permeable FMK Caspase Probes
- Prepare a 2-5 mM stock solution in DMSO.
- Treat cells as desired.
- Prepare a 2X permeable caspase substrate (20 µM) assay solution by diluting the DMSO stock solution (from Step 2.1) in Hanks with 20 mM Hepes buffer (HHBS).
- Mix equal volume of the treated cells with 2X caspase substrate assay solution (from Step 2.3), and incubate the cells in a 37°C, 5% CO2 incubator for at least1 hour.
- Wash the cells with HHBS for at least once.
- Monitor the fluorescence intensity by a flow cytometer, a fluorescence microscope or a fluorescence microplate reader.
Cell Caspase Assays Using Cell-Permeable FMK Caspase Probes (For #13470-13476 only)
- Prepare a 250X stock solution by adding 50 µL DMSO into the vial.
- Treat cells as desired.
- Add 250 X DMSO stock solution into the cell solution at a 1:250 ratio (such as 2 µL to 500 µL cells), and incubate the cells in a 37°C, 5% CO2 incubator for 1 hour.
- Wash the cells with HHBS for at least once.
- Monitor the fluorescence intensity by flow cytometer, fluorescence microscopy or fluorescent microplate reader.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of TF3-DEVD-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 | 102.56 µL | 512.799 µL | 1.026 mL | 5.128 mL | 10.256 mL |
| 5 mM | 20.512 µL | 102.56 µL | 205.12 µL | 1.026 mL | 2.051 mL |
| 10 mM | 10.256 µL | 51.28 µL | 102.56 µL | 512.799 µL | 1.026 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
| Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
| / | = | x | = |
Spectrum
Open in Advanced Spectrum Viewer
Spectral properties
| Correction Factor (280 nm) | 0.179 |
| Extinction coefficient (cm -1 M -1) | 750001 |
| Excitation (nm) | 554 |
| Emission (nm) | 578 |
Images
Figure 1. Chemical structure for TF3-DEVD-FMK.
Citations
View all 29 citations: Citation Explorer
The Dietary Flavonoid Fisetin Causes Cell Cycle Arrest, Caspase-Dependent Apoptosis, and Enhanced Cytotoxicity of Chemotherapeutic Drugs in Triple-Negative Breast Cancer Cells
Authors: Smith, M. L., Murphy, K., Doucette, C. D., Greenshields, A. L., Hoskin, D. W.
Journal: J Cell Biochem (2016): 1913-25
Authors: Smith, M. L., Murphy, K., Doucette, C. D., Greenshields, A. L., Hoskin, D. W.
Journal: J Cell Biochem (2016): 1913-25
Contribution of membrane progesterone receptor alpha to the induction of progesterone-mediated apoptosis associated with mitochondrial membrane disruption and caspase cascade activation in Jurkat cell lines
Authors: Kon, A., Yuan, B., Hanazawa, T., Kikuchi, H., Sato, M., Furutani, R., Takagi, N., Toyoda, H.
Journal: Oncol Rep (2013): 1965-70
Authors: Kon, A., Yuan, B., Hanazawa, T., Kikuchi, H., Sato, M., Furutani, R., Takagi, N., Toyoda, H.
Journal: Oncol Rep (2013): 1965-70
TNF induces caspase-dependent inflammation in renal endothelial cells through a Rho- and myosin light chain kinase-dependent mechanism
Authors: Wu, X., Guo, R., Chen, P., Wang, Q., Cunningham, P. N.
Journal: Am J Physiol Renal Physiol (2009): F316-26
Authors: Wu, X., Guo, R., Chen, P., Wang, Q., Cunningham, P. N.
Journal: Am J Physiol Renal Physiol (2009): F316-26
Pan-caspase inhibition suppresses polyethylene particle-induced osteolysis
Authors: L, undefined and graeber, S., Jaeckel, S., Loer, F., Wedemeyer, C., Hilken, G., Canbay, A., Totsch, M., von Knoch, M.
Journal: Apoptosis (2009): 173-81
Authors: L, undefined and graeber, S., Jaeckel, S., Loer, F., Wedemeyer, C., Hilken, G., Canbay, A., Totsch, M., von Knoch, M.
Journal: Apoptosis (2009): 173-81
Inhibition of Caspase-Like Activities Prevents the Appearance of Reactive Oxygen Species and Dark-Induced Apoptosis in the Unicellular Chlorophyte Dunaliella Tertiolecta(1)
Authors: Segovia, M., Berges, J. A.
Journal: J Phycol (2009): 1116-26
Authors: Segovia, M., Berges, J. A.
Journal: J Phycol (2009): 1116-26
Silvestrol, a potential anticancer rocaglate derivative from Aglaia foveolata, induces apoptosis in LNCaP cells through the mitochondrial/apoptosome pathway without activation of executioner caspase-3 or -7
Authors: Kim, S., Hwang, B. Y., Su, B. N., Chai, H., Mi, Q., Kinghorn, A. D., Wild, R., Swanson, S. M.
Journal: Anticancer Res (2007): 2175-83
Authors: Kim, S., Hwang, B. Y., Su, B. N., Chai, H., Mi, Q., Kinghorn, A. D., Wild, R., Swanson, S. M.
Journal: Anticancer Res (2007): 2175-83
Leptin induces interleukin-1beta release from rat microglial cells through a caspase 1 independent mechanism
Authors: Pinteaux, E., Inoue, W., Schmidt, L., Molina-Holgado, F., Rothwell, N. J., Luheshi, G. N.
Journal: J Neurochem (2007): 826-33
Authors: Pinteaux, E., Inoue, W., Schmidt, L., Molina-Holgado, F., Rothwell, N. J., Luheshi, G. N.
Journal: J Neurochem (2007): 826-33
Lipopolysaccharide accelerates caspase-independent but cathepsin B-dependent death of human lung epithelial cells
Authors: Tang, P. S., Tsang, M. E., Lodyga, M., Bai, X. H., Miller, A., Han, B., Liu, M.
Journal: J Cell Physiol (2006): 457-67
Authors: Tang, P. S., Tsang, M. E., Lodyga, M., Bai, X. H., Miller, A., Han, B., Liu, M.
Journal: J Cell Physiol (2006): 457-67
Selective caspase activation may contribute to neurological dysfunction after experimental spinal cord trauma
Authors: Knoblach, S. M., Huang, X., VanGelderen, J., Calva-Cerqueira, D., Faden, A. I.
Journal: J Neurosci Res (2005): 369-80
Authors: Knoblach, S. M., Huang, X., VanGelderen, J., Calva-Cerqueira, D., Faden, A. I.
Journal: J Neurosci Res (2005): 369-80
C-terminal region of Bfl-1 induces cell death that accompanies caspase activation when fused with GFP
Authors: Yang, W. S., Ko, J. K., Park, S. O., Choi, H. Y., Kim, Y. N., Kim, C. W.
Journal: J Cell Biochem (2005): 1234-47
Authors: Yang, W. S., Ko, J. K., Park, S. O., Choi, H. Y., Kim, Y. N., Kim, C. W.
Journal: J Cell Biochem (2005): 1234-47
References
View all 26 references: Citation Explorer
Suppression of human T cell proliferation by the caspase inhibitors, z-VAD-FMK and z-IETD-FMK is independent of their caspase inhibition properties
Authors: Lawrence CP, Chow SC.
Journal: Toxicol Appl Pharmacol. (2012)
Authors: Lawrence CP, Chow SC.
Journal: Toxicol Appl Pharmacol. (2012)
Inhibition of elicitation of allergic contact dermatitis by topical use of Z-VAD-FMK, a broad caspase inhibitor: experiment in mice
Authors: Li YY, Yan CL.
Journal: Zhonghua Yi Xue Za Zhi (2012): 1992
Authors: Li YY, Yan CL.
Journal: Zhonghua Yi Xue Za Zhi (2012): 1992
Structure of human caspase-6 in complex with Z-VAD-FMK: New peptide binding mode observed for the non-canonical caspase conformation
Authors: Muller I, Lamers MB, Ritchie AJ, Dominguez C, Munoz-Sanjuan I, Kiselyov A.
Journal: Bioorg Med Chem Lett (2011): 5244
Authors: Muller I, Lamers MB, Ritchie AJ, Dominguez C, Munoz-Sanjuan I, Kiselyov A.
Journal: Bioorg Med Chem Lett (2011): 5244
Intracochlear perfusion of leupeptin and z-VAD-FMK: influence of antiapoptotic agents on gunshot-induced hearing loss
Authors: Abaamrane L, Raffin F, Schmerber S, Sendowski I.
Journal: Eur Arch Otorhinolaryngol (2011): 987
Authors: Abaamrane L, Raffin F, Schmerber S, Sendowski I.
Journal: Eur Arch Otorhinolaryngol (2011): 987
Plasmodium falciparum metacaspase PfMCA-1 triggers a z-VAD-fmk inhibitable protease to promote cell death
Authors: Meslin B, Beavogui AH, Fasel N, Picot S.
Journal: PLoS One (2011): e23867
Authors: Meslin B, Beavogui AH, Fasel N, Picot S.
Journal: PLoS One (2011): e23867
Pretreatment with pancaspase inhibitor (Z-VAD-FMK) delays but does not prevent intraperitoneal heat-killed group B Streptococcus-induced preterm delivery in a pregnant mouse model
Authors: Equils O, Moffatt-Blue C, Ishikawa TO, Simmons CF, Ilievski V, Hirsch E.
Journal: Infect Dis Obstet Gynecol (2009): 749432
Authors: Equils O, Moffatt-Blue C, Ishikawa TO, Simmons CF, Ilievski V, Hirsch E.
Journal: Infect Dis Obstet Gynecol (2009): 749432
Attenuation of allergic contact dermatitis by Z-VAD-FMK, a broad caspase inhibitor: experiment with mice
Authors: Li YY, Yan CL, Xu W.
Journal: Zhonghua Yi Xue Za Zhi (2008): 3153
Authors: Li YY, Yan CL, Xu W.
Journal: Zhonghua Yi Xue Za Zhi (2008): 3153
Bodipy-VAD-Fmk, a useful tool to study yeast peptide N-glycanase activity
Authors: Witte MD, Descals CV, de Lavoir SV, Florea BI, van der Marel GA, Overkleeft HS.
Journal: Org Biomol Chem (2007): 3690
Authors: Witte MD, Descals CV, de Lavoir SV, Florea BI, van der Marel GA, Overkleeft HS.
Journal: Org Biomol Chem (2007): 3690
Effects of caspase inhibitors (z-VAD-fmk, z-VDVAD-fmk) on Nile Red fluorescence pattern in 7-ketocholesterol-treated cells: investigation by flow cytometry and spectral imaging microscopy
Authors: Vejux A, Lizard G, Tourneur Y, Riedinger JM, Frouin F, Kahn E.
Journal: Cytometry A (2007): 550
Authors: Vejux A, Lizard G, Tourneur Y, Riedinger JM, Frouin F, Kahn E.
Journal: Cytometry A (2007): 550
Caspase inhibitor Z-VAD-FMK enhances the freeze-thaw survival rate of human embryonic stem cells
Authors: Heng BC, Clement MV, Cao T.
Journal: Biosci Rep (2007): 257
Authors: Heng BC, Clement MV, Cao T.
Journal: Biosci Rep (2007): 257
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?
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