FITC-C6-LEHD-FMK

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
Quotation	Request
International	See distributors
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	1031.07
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

1031.07

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-LEHD-FMK provides a convenient means for sensitive detection of activated caspase-9 in living cells. FITC-LEHD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-9 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-LEHD-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	96.987 µL	484.933 µL	969.866 µL	4.849 mL	9.699 mL
5 mM	19.397 µL	96.987 µL	193.973 µL	969.866 µL	1.94 mL
10 mM	9.699 µL	48.493 µL	96.987 µL	484.933 µL	969.866 µL

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-DEVD-FMK	491	516	73000	0.92	0.35

Images

Figure 1. Chemical structure for FITC-C6-LEHD-FMK

References

View all 9 references: Citation Explorer

Endoplasmic reticulum stress and trophic factor withdrawal activate distinct signaling cascades that induce glycogen synthase kinase-3 beta and a caspase-9-dependent apoptosis in cerebellar granule neurons
Authors: Brewster JL, Linseman DA, Bouchard RJ, Loucks FA, Precht TA, Esch EA, Heidenreich KA.
Journal: Mol Cell Neurosci (2006): 242

Enhanced 15-HPETE production during oxidant stress induces apoptosis of endothelial cells
Authors: Sordillo LM, Weaver JA, Cao YZ, Corl C, Sylte MJ, Mullarky IK.
Journal: Prostaglandins Other Lipid Mediat (2005): 19

P2X7 receptor-mediated apoptosis of human cervical epithelial cells
Authors: Wang Q, Wang L, Feng YH, Li X, Zeng R, Gorodeski GI.
Journal: Am J Physiol Cell Physiol (2004): C1349

Caspase activation during Haemophilus somnus lipooligosaccharide-mediated apoptosis of bovine endothelial cells
Authors: Sylte MJ, Leite FP, Kuckleburg CJ, Inzana TJ, Czuprynski CJ.
Journal: Microb Pathog (2003): 285

Induction of apoptosis by apicidin, a histone deacetylase inhibitor, via the activation of mitochondria-dependent caspase cascades in human Bcr-Abl-positive leukemia cells
Authors: Cheong JW, Chong SY, Kim JY, Eom JI, Jeung HK, Maeng HY, Lee ST, Min YH.
Journal: Clin Cancer Res (2003): 5018

Protein kinase Cdelta amplifies ceramide formation via mitochondrial signaling in prostate cancer cells
Authors: Sumitomo M, Ohba M, Asakuma J, Asano T, Kuroki T, Hayakawa M.
Journal: J Clin Invest (2002): 827

3-m-bromoacetylamino benzoic acid ethyl ester: a new cancericidal agent that activates the apoptotic pathway through caspase-9
Authors: Schlesinger M, Jiang JD, Roboz JP, Denner L, Ling YH, Holl and JF, Bekesi JG.
Journal: Biochem Pharmacol (2000): 1693

Human malignant glioma therapy using anti-alpha(v)beta3 integrin agents
Authors: Chatterjee S, Matsumura A, Schradermeier J, Gillespie GY.
Journal: J Neurooncol (2000): 135

Phenylephrine protects neonatal rat cardiomyocytes from hypoxia and serum deprivation-induced apoptosis
Authors: Zhu H, McElwee-Witmer S, Perrone M, Clark KL, Zilberstein A.
Journal: Cell Death Differ (2000): 773