Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>

FITC-C6-DEVD-FMK

Chemical structure for FITC-C6-DEVD-FMK
Chemical structure for FITC-C6-DEVD-FMK
Ordering information
Price ()
Catalog Number13406
Unit Size
Find Distributor
Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Physical properties
Molecular weight994.99
SolventDMSO
Spectral properties
Correction Factor (280 nm)0.254
Extinction coefficient (cm -1 M -1)73000
Excitation (nm)491
Emission (nm)516
Quantum yield0.92
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


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

  1. Prepare a 10 mM stock solution in DMSO.

  2. 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.

  3. 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.

  4. Monitor the fluorescence using a fluorescence microplate reader, or absorbance using an absorbance microplate reader.

Cell Caspase Assays Using Cell-Permeable FMK Caspase Probes

  1. Prepare a 2-5 mM stock solution in DMSO.

  2. Treat cells as desired.

  3. 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).

  4. 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.

  5. Wash the cells with HHBS for at least once.

  6. 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)

  1. Prepare a 250X stock solution by adding 50 µL DMSO into the vial.

  2. Treat cells as desired.

  3. 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.

  4. Wash the cells with HHBS for at least once.

  5. 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 mg0.5 mg1 mg5 mg10 mg
1 mM100.504 µL502.518 µL1.005 mL5.025 mL10.05 mL
5 mM20.101 µL100.504 µL201.007 µL1.005 mL2.01 mL
10 mM10.05 µL50.252 µL100.504 µL502.518 µL1.005 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
/=x=

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Correction Factor (280 nm)0.254
Extinction coefficient (cm -1 M -1)73000
Excitation (nm)491
Emission (nm)516
Quantum yield0.92

Product family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (280 nm)
FITC-C6-LEHD-FMK491516730000.920.254

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)

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
Antitumor mechanism of 3-bromopropionylamino benzoylurea on leukemia and lymphoma
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
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