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Ac-DEVD-AMC *CAS 169332-61-0*

Ac-DEVD-AMC is a fluorogenic substrate for caspase 3, a protease that is rapidly activated when cells are exposed to apoptotic conditions and that cleaves poly(ADP-ribose) polymerase.

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-DEVD-AMC *CAS 169332-61-0* 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 mM148.008 µL740.039 µL1.48 mL7.4 mL14.801 mL
5 mM29.602 µL148.008 µL296.016 µL1.48 mL2.96 mL
10 mM14.801 µL74.004 µL148.008 µL740.039 µL1.48 mL

Molarity calculator

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Spectrum

Product family

Citations

View all 5 citations: Citation Explorer
Assessment of caspase activity in post mortem muscle as a way to explain characteristics of DFD beef
Authors: Fuente-Garc{\'\i}a, Claudia and Aldai, Noelia and Sentandreu, Enrique and Oliv{\'a}n, Mamen and Franco, Daniel and Garc{\'\i}a-Torres, Susana and Sentandreu, Miguel {\'A}ngel
Journal: Journal of Food Composition and Analysis (2022): 104599
Ipatasertib, an oral AKT inhibitor, inhibits cell proliferation and migration, and induces apoptosis in serous endometrial cancer
Authors: Buckingham, Lindsey and Hao, Tianran and O’Donnell, Jillian and Zhao, Ziyi and Zhang, Xin and Fan, Yali and Sun, Wenchuan and Zhang, Yingao and Suo, Hongyan and Secord, Angeles Alvarez and others,
Journal: American Journal of Cancer Research (2022): 2850
ONC206, an imipridone derivative, induces cell death through activation of the integrated stress response in serous endometrial cancer in vitro
Authors: Zhang, Yingao and Huang, Yu and Yin, Yajie and Fan, Yali and Sun, Wenchuan and Zhao, Xiaoling and Tucker, Katherine and Staley, Allison and Paraghamian, Sarah and Hawkins, Gabrielle and others,
Journal: Frontiers in Oncology (2020): 577141
Overexpression of ILK promotes temozolomide resistance in glioma cells
Authors: Liang, Feng and Wang, Bing and Bao, Long and Zhao, Yin-Sheng and Zhang, Shi-Min and Zhang, Shu-Qin
Journal: Molecular medicine reports (2017): 1297--1304
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
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
Serofendic acid, a neuroprotective substance derived from fetal calf serum, inhibits mitochondrial membrane depolarization and caspase-3 activation
Authors: Kume T, Taguchi R, Katsuki H, Akao M, Sugimoto H, Kaneko S, Akaike A.
Journal: Eur J Pharmacol (2006): 69
Multiparameter measurement of caspase 3 activation and apoptotic cell death in NT2 neuronal precursor cells using high-content analysis
Authors: Fennell M, Chan H, Wood A.
Journal: J Biomol Screen (2006): 296
Asymmetric dimethylarginine induces apoptosis via p38 MAPK/caspase-3-dependent signaling pathway in endothelial cells
Authors: Jiang DJ, Jia SJ, Dai Z, Li YJ.
Journal: J Mol Cell Cardiol (2006): 529
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 December 6, 2024

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Physical properties

Molecular weight

675.64

Solvent

DMSO

Spectral properties

Excitation (nm)

341

Emission (nm)

441

Storage, safety and handling

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

Storage

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

CAS

169332-61-0
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