FITC-C6-DEVD-FMK
| Overview |  SDSProtocol |
Molecular weight 994.99 | Correction Factor (280 nm) 0.254 | Extinction coefficient (cm -1 M -1) 73000 | Excitation (nm) 491 | Emission (nm) 516 | Quantum yield 0.92 |
Activation of caspases plays a central role in apoptosis. FITC-C6-DEVD-FMK provides a convenient means for sensitive detection of activated caspase-3 in living cells. FITC-DEVD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-3 in apoptotic cells. The FITC label allows for direct detection of activated caspases in apoptotic cells by fluorescence microscopy, flow cytometry, or fluorescence plate 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 FITC-C6-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 | 100.504 µL | 502.518 µL | 1.005 mL | 5.025 mL | 10.05 mL |
| 5 mM | 20.101 µL | 100.504 µL | 201.007 µL | 1.005 mL | 2.01 mL |
| 10 mM | 10.05 µL | 50.252 µL | 100.504 µL | 502.518 µL | 1.005 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
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Spectral properties
| Correction Factor (280 nm) | 0.254 |
| Extinction coefficient (cm -1 M -1) | 73000 |
| Excitation (nm) | 491 |
| Emission (nm) | 516 |
| Quantum yield | 0.92 |
Product family
| Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (280 nm) |
| FITC-C6-LEHD-FMK | 491 | 516 | 73000 | 0.92 | 0.254 |
Images
Figure 1. Chemical structure for FITC-C6-DEVD-FMK
Citations
View all 1 citations: Citation Explorer
The BAD-BAX-caspase-3 cascade modulates synaptic vesicle pools via autophagy
Authors: Gu, Qinhua and Jiao, Song and Duan, Kaizheng and Wang, Ya-Xian and Petralia, Ronald S and Li, Zheng
Journal: Journal of Neuroscience (2020)
Authors: Gu, Qinhua and Jiao, Song and Duan, Kaizheng and Wang, Ya-Xian and Petralia, Ronald S and Li, Zheng
Journal: Journal of Neuroscience (2020)
References
View all 4 references: Citation Explorer
Activation of caspase-like activity and poly (ADP-ribose) polymerase degradation during sporulation in Aspergillus nidulans
Authors: Thrane C, Kaufmann U, Stummann BM, Olsson S.
Journal: Fungal Genet Biol (2004): 361
Authors: Thrane C, Kaufmann U, Stummann BM, Olsson S.
Journal: Fungal Genet Biol (2004): 361
Antitumor mechanism of 3-bromopropionylamino benzoylurea on leukemia and lymphoma
Authors: Li JN, Song DQ, Jiang JD.
Journal: Yao Xue Xue Bao (2004): 491
Authors: Li JN, Song DQ, Jiang JD.
Journal: Yao Xue Xue Bao (2004): 491
Caspase-dependent and independent cell death in rat hepatoma 5123tc cells
Authors: P, undefined and ey S, Smith B, Walker PR, Sikorska M.
Journal: Apoptosis (2000): 265
Authors: P, undefined and ey S, Smith B, Walker PR, Sikorska M.
Journal: Apoptosis (2000): 265
C-myc antisense oligodeoxynucleotides can induce apoptosis and down-regulate Fas expression in rheumatoid synoviocytes
Authors: Hashiramoto A, Sano H, Maekawa T, Kawahito Y, Kimura S, Kusaka Y, Wilder RL, Kato H, Kondo M, Nakajima H.
Journal: Arthritis Rheum (1999): 954
Authors: Hashiramoto A, Sano H, Maekawa T, Kawahito Y, Kimura S, Kusaka Y, Wilder RL, Kato H, Kondo M, Nakajima H.
Journal: Arthritis Rheum (1999): 954