FITC-C6-DEVD-FMK
Overview | ![]() ![]() |
See also: Apoptosis and Necrosis, Caspases, Cell Viability Assays, Cellular Processes, Chemical Reagents, Classic Dyes, Rhodamines and Rhodamine Derivatives, Enzymes, Fluoresceins
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 |
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
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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