Ac-DEVD-AFC *CAS 201608-14-2*
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See also: Apoptosis and Necrosis, Caspases, Cell Viability Assays, Cellular Processes, Chemical Reagents, Classic Dyes, Coumarins, Enzymes
CAS 201608-14-2 | Molecular weight 729.61 | Extinction coefficient (cm -1 M -1) 170001 | Excitation (nm) 376 | Emission (nm) 482 | Quantum yield 0.531 |
Ac-DEVD-AFC is a fluorogenic substrate for caspase 3, a protease that is rapidly activated when cells are exposed to apoptotic conditions and that cleaves poly(ADP-ribose) polymerase. The 7-amido-4-trifluoromethylcoumarin derivatives have better membrane permeability than the 7-amido-4-methylcoumarin derivatives.
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 Ac-DEVD-AFC *CAS 201608-14-2* 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 | 137.06 µL | 685.298 µL | 1.371 mL | 6.853 mL | 13.706 mL |
5 mM | 27.412 µL | 137.06 µL | 274.119 µL | 1.371 mL | 2.741 mL |
10 mM | 13.706 µL | 68.53 µL | 137.06 µL | 685.298 µL | 1.371 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Open in Advanced Spectrum Viewer
Spectral properties
Extinction coefficient (cm -1 M -1) | 170001 |
Excitation (nm) | 376 |
Emission (nm) | 482 |
Quantum yield | 0.531 |
Product family
Name | Excitation (nm) | Emission (nm) |
Ac-DEVD-AMC *CAS 169332-61-0* | 341 | 441 |
Ac-DEVD-CHO *CAS 169332-60-9* | - | - |
Ac-DEVD-pNA *CAS 189950-66-1* | - | - |
Ac-IETD-AFC *CAS 211990-57-7* | 376 | 482 |
Z-DEVD-AFC | 376 | 482 |
Citations
View all 5 citations: Citation Explorer
Mitophagy regulates mitochondrial network signaling, oxidative stress, and apoptosis during myoblast differentiation
Authors: Baechler, Brittany L and Bloemberg, Darin and Quadrilatero, Joe
Journal: Autophagy (2019): 1606--1619
Authors: Baechler, Brittany L and Bloemberg, Darin and Quadrilatero, Joe
Journal: Autophagy (2019): 1606--1619
Regulation of glioblastoma cell proliferation in dependence of cell density and growth factors in vitro
Authors: Liu, Yun
Journal: (2019)
Authors: Liu, Yun
Journal: (2019)
SOX9 in the development and chemotherapy of cholangiocarcinoma (CCA)
Authors: Yuan, Xiaodong
Journal: (2018)
Authors: Yuan, Xiaodong
Journal: (2018)
Structure-dependent induction of apoptosis by hepatotoxic pyrrolizidine alkaloids in the human hepatoma cell line HepaRG: Single versus repeated exposure
Authors: Waizenegger, Julia and Braeuning, Albert and Templin, Markus and Lampen, Alfonso and Hessel-Pras, Stefanie
Journal: Food and Chemical Toxicology (2018)
Authors: Waizenegger, Julia and Braeuning, Albert and Templin, Markus and Lampen, Alfonso and Hessel-Pras, Stefanie
Journal: Food and Chemical Toxicology (2018)
pH-Assisted surface functionalization of selenium nanoparticles with curcumin to achieve enhanced cancer chemopreventive activity
Authors: Yu, Shaoxuan and Wang, Yanru and Zhang, Wentao and Zhang, Yuhuan and Zhu, Wenxin and Liu, Yingnan and Zhang, Daohong and Wang, Jianlong
Journal: RSC Advances (2016): 72213--72223
Authors: Yu, Shaoxuan and Wang, Yanru and Zhang, Wentao and Zhang, Yuhuan and Zhu, Wenxin and Liu, Yingnan and Zhang, Daohong and Wang, Jianlong
Journal: RSC Advances (2016): 72213--72223
References
View all 67 references: Citation Explorer
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Measurement of two caspase activities simultaneously in living cells by a novel dual FRET fluorescent indicator probe
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Journal: Cytometry A (2006): 477
Quantitative measurement of caspase-3 activity in a living starfish egg
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Photoreceptor cell apoptosis induced by the 2-nitroimidazole radiosensitizer, CI-1010, is mediated by p53-linked activation of caspase-3
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Diallyl Trisulfide Induces Apoptosis of Human Gastric Cancer Cell Line MGC803 Through Caspase-3 Pathway.
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Asymmetric dimethylarginine induces apoptosis via p38 MAPK/caspase-3-dependent signaling pathway in endothelial cells
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Multiparameter measurement of caspase 3 activation and apoptotic cell death in NT2 neuronal precursor cells using high-content analysis
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Serofendic acid, a neuroprotective substance derived from fetal calf serum, inhibits mitochondrial membrane depolarization and caspase-3 activation
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Application notes
Abbreviation of Common Chemical Compounds Related to Peptides
Abbreviation of Common Chemical Compounds Related to Peptides
Abbreviation of Common Chemical Compounds Related to Peptides
Abbreviation of Common Chemical Compounds Related to Peptides
Abbreviation of Common Chemical Compounds Related to Peptides
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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