FITC-C6-DEVD-FMK

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Physical properties

Molecular weight	994.99
Solvent	DMSO

Spectral properties

Correction Factor (280 nm)	0.35
Extinction coefficient (cm ^-1 M ^-1)	73000
Excitation (nm)	491
Emission (nm)	516
Quantum yield	0.92

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200

Overview SDS Protocol

Molecular weight

994.99

Correction Factor (280 nm)

0.35

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
	/		=		x		=

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Correction Factor (280 nm)	0.35
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	0.35

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)

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