logo
AAT Bioquest

Ac-IETD-AFC *CAS 211990-57-7*

Ac-IETD-AFC is a fluorogenic caspase-8/granzyme B substrate containing the acetyl (Ac) moiety. This substrate is hydrlyzed by caspase 8 to generate highly fluorescent 7-amido-4-trifluoromethylcoumarin (AFC).

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 Ac-IETD-AFC *CAS 211990-57-7* 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 mM137.052 µL685.26 µL1.371 mL6.853 mL13.705 mL
5 mM27.41 µL137.052 µL274.104 µL1.371 mL2.741 mL
10 mM13.705 µL68.526 µL137.052 µL685.26 µL1.371 mL

Molarity calculator

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

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

Spectrum

Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yield
Ac-DEVD-AFC *CAS 201608-14-2*3764821700010.531
Ac-IETD-AMC341441--
Z-IETD-AFC *CAS 219138-02-0*3764821700010.531

Citations

View all 1 citations: Citation Explorer
pH-Assisted surface functionalization of selenium nanoparticles with curcumin to achieve enhanced cancer chemopreventive activity
Authors: Yu, Shaoxuan and Wang, Yanru and Zhang, Wentao and Zhang, Yuhuan and Zhu, Wenxin and Liu, Yingnan and Zhang, Daohong and Wang, Jianlong
Journal: RSC Advances (2016): 72213--72223

References

View all 67 references: Citation Explorer
In vivo and in vitro sensitization of leukemic cells to adriamycin-induced apoptosis by pentoxifylline. Involvement of caspase cascades and IkappaBalpha phosphorylation
Authors: Lerma-Diaz JM, Hern and ez-Flores G, Dominguez-Rodriguez JR, Ortiz-Lazareno PC, Gomez-Contreras P, Cervantes-Munguia R, Scott-Algara D, Aguilar-Lemarroy A, Jave-Suarez LF, Bravo-Cuellar A.
Journal: Immunol Lett (2006): 149
Measurement of two caspase activities simultaneously in living cells by a novel dual FRET fluorescent indicator probe
Authors: Wu X, Simone J, Hewgill D, Siegel R, Lipsky PE, He L.
Journal: Cytometry A (2006): 477
Quantitative measurement of caspase-3 activity in a living starfish egg
Authors: Sakaue M, Motoyama Y, Yamamoto K, Shiba T, Teshima T, Chiba K.
Journal: Biochem Biophys Res Commun (2006): 878
Photoreceptor cell apoptosis induced by the 2-nitroimidazole radiosensitizer, CI-1010, is mediated by p53-linked activation of caspase-3
Authors: Miller TJ, Schneider RJ, Miller JA, Martin BP, Al-Ubaidi MR, Agarwal N, Dethloff LA, Philbert MA.
Journal: Neurotoxicology (2006): 44
Diallyl Trisulfide Induces Apoptosis of Human Gastric Cancer Cell Line MGC803 Through Caspase-3 Pathway.
Authors: Xiao XL, Peng J, Su Q, Xiang SL, Tang GH, Huang YS, Zhou XT.
Journal: Ai Zheng (2006): 1247
Page updated on October 10, 2024

Ordering information

Price
Unit size
Catalog Number13410
Quantity
Add to cart

Additional ordering information

Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
Technical SupportContact us
Purchase orderSend to sales@aatbio.com
ShippingStandard overnight for United States, inquire for international
Request quotation

Physical properties

Molecular weight

729.65

Solvent

DMSO

Spectral properties

Extinction coefficient (cm -1 M -1)

170001

Excitation (nm)

376

Emission (nm)

482

Quantum yield

0.531

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200

CAS

211990-57-7
Product Image
Product Image
Gallery Image 1