Z-IETD-ProRed™ 620
Ordering information
Price | |
Catalog Number | |
Unit Size | |
Quantity |
Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | 1565.59 |
Solvent | DMSO |
Spectral properties
Excitation (nm) | 532 |
Emission (nm) | 619 |
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 | ![]() ![]() |
See also: Caspases
Molecular weight 1565.59 | Excitation (nm) 532 | Emission (nm) 619 |
ProRed™-derived protease substrates are colorless and non-fluorescent. Cleavage of blocking protease-cleavable peptide residue by caspases generates the strongly red fluorescent ProRed™ that can be monitored fluorimetrically at ~620 nm with excitation of ~530 nm. ProRed™-derived caspase substrates are the most sensitive red indicators for the fluorimetric detection of various caspase activities. This IETD-ProRed™ substrate is specific for detecting caspase 8.
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 Z-IETD-ProRed™ 620 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 | 63.874 µL | 319.368 µL | 638.737 µL | 3.194 mL | 6.387 mL |
5 mM | 12.775 µL | 63.874 µL | 127.747 µL | 638.737 µL | 1.277 mL |
10 mM | 6.387 µL | 31.937 µL | 63.874 µL | 319.368 µL | 638.737 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Images

Figure 1. Detection of Caspase 8 Activity in Jurkat cells with Z-IETD-ProRed™ 620. Jurkat cells were seeded on the same day at 200,000 cells/90 µL/well in a Costar black wall/clear bottom 96-well plate. The cells were treated with staurosporine at the final concentration of 1 µM for 5 hours while the untreated cells were used as control. The caspase 8 assay solution (100 µL/well) was added and incubated at room temperature for 1 hour. The fluorescence intensity was measured at Ex/Em = 540/620 nm with a FlexStation™ microplate reader (Molecular Devices).
Citations
View all 1 citations: Citation Explorer
Degenerin channel activation causes caspase-mediated protein degradation and mitochondrial dysfunction in adult C. elegans muscle
Authors: Gaffney, Christopher J and Shephard, Freya and Chu, Jeff and Baillie, David L and Rose, Ann and Constantin-Teodosiu, Dumitru and Greenhaff, Paul L and Szewczyk, Nathaniel J
Journal: Journal of Cachexia, Sarcopenia and Muscle (2015)
Authors: Gaffney, Christopher J and Shephard, Freya and Chu, Jeff and Baillie, David L and Rose, Ann and Constantin-Teodosiu, Dumitru and Greenhaff, Paul L and Szewczyk, Nathaniel J
Journal: Journal of Cachexia, Sarcopenia and Muscle (2015)
References
View all 101 references: Citation Explorer
3,5-dibenzyloxy-4'-hydroxystilbene induces early caspase-9 activation during apoptosis in human K562 chronic myelogenous leukemia cells
Authors: Roslie H, Chan KM, Rajab NF, Velu SS, Kadir SA, Bunyamin I, Weber JF, Thomas NF, Majeed AB, Myatt G, Inayat-Hussain SH.
Journal: J Toxicol Sci (2012): 13
Authors: Roslie H, Chan KM, Rajab NF, Velu SS, Kadir SA, Bunyamin I, Weber JF, Thomas NF, Majeed AB, Myatt G, Inayat-Hussain SH.
Journal: J Toxicol Sci (2012): 13
Dopaminochrome induces caspase-independent apoptosis in the mesencephalic cell line, MN9D
Authors: Linsenbardt AJ, Breckenridge JM, Wilken GH, Macarthur H.
Journal: J Neurochem. (2012)
Authors: Linsenbardt AJ, Breckenridge JM, Wilken GH, Macarthur H.
Journal: J Neurochem. (2012)
Acacetin induces apoptosis in human T cell leukemia Jurkat cells via activation of a caspase cascade
Authors: Watanabe K, Kanno S, Tomizawa A, Yomogida S, Ishikawa M.
Journal: Oncol Rep (2012): 204
Authors: Watanabe K, Kanno S, Tomizawa A, Yomogida S, Ishikawa M.
Journal: Oncol Rep (2012): 204
5-Phenylselenyl- and 5-methylselenyl-methyl-2'-deoxyuridine induce oxidative stress, DNA damage, and caspase-2-dependent apoptosis in cancer cells
Authors: Kim BM, Rode AB, Han EJ, Hong IS, Hong SH.
Journal: Apoptosis (2012): 200
Authors: Kim BM, Rode AB, Han EJ, Hong IS, Hong SH.
Journal: Apoptosis (2012): 200
High cytosolic free calcium level signals apoptosis through mitochondria-caspase mediated pathway in rat eggs cultured in vitro
Authors: Tripathi A, Chaube SK.
Journal: Apoptosis (2012): 439
Authors: Tripathi A, Chaube SK.
Journal: Apoptosis (2012): 439
The effect of low-level laser to apoptosis of chondrocyte and caspases expression, including caspase-8 and caspase-3 in rabbit surgery-induced model of knee osteoarthritis
Authors: Lin HD, He CQ, Luo QL, Zhang JL, Zeng DX.
Journal: Rheumatol Int (2012): 759
Authors: Lin HD, He CQ, Luo QL, Zhang JL, Zeng DX.
Journal: Rheumatol Int (2012): 759
A caspase 8-based suicide switch induces apoptosis in nanobody-directed chimeric receptor expressing T cells
Authors: Khaleghi S, Rahbarizadeh F, Ahmadv and D, Rasaee MJ, Pognonec P.
Journal: Int J Hematol (2012): 434
Authors: Khaleghi S, Rahbarizadeh F, Ahmadv and D, Rasaee MJ, Pognonec P.
Journal: Int J Hematol (2012): 434
Taurine protects rat testes against doxorubicin-induced oxidative stress as well as p53, Fas and caspase 12-mediated apoptosis
Authors: Das J, Ghosh J, Manna P, Sil PC.
Journal: Amino Acids (2012): 1839
Authors: Das J, Ghosh J, Manna P, Sil PC.
Journal: Amino Acids (2012): 1839
Low-Dose Spironolactone Prevents Apoptosis Repressor With Caspase Recruitment Domain Degradation During Myocardial Infarction
Authors: Loan Le TY, Mardini M, Howell VM, Funder JW, Ashton AW, Mihailidou AS.
Journal: Hypertension. (2012)
Authors: Loan Le TY, Mardini M, Howell VM, Funder JW, Ashton AW, Mihailidou AS.
Journal: Hypertension. (2012)
Steroid Receptor Coactivator-interacting Protein (SIP) Inhibits Caspase-independent Apoptosis by Preventing Apoptosis-inducing Factor (AIF) from Being Released from Mitochondria
Authors: Wang D, Liang J, Zhang Y, Gui B, Wang F, Yi X, Sun L, Yao Z, Shang Y.
Journal: J Biol Chem (2012): 12612
Authors: Wang D, Liang J, Zhang Y, Gui B, Wang F, Yi X, Sun L, Yao Z, Shang Y.
Journal: J Biol Chem (2012): 12612
Application notes
FAQ
Are inflammasomes and caspase-1 related?
Do you offer any fluorimetric assays that measure caspase activation/activity in live cells using a flow cytometer?
Does pH and staining temperature affect Annexin V-Phosphatidylserine binding?
Does propidium iodide stain apoptotic cells?
How can I tell if my cell sample is dying?
Do you offer any fluorimetric assays that measure caspase activation/activity in live cells using a flow cytometer?
Does pH and staining temperature affect Annexin V-Phosphatidylserine binding?
Does propidium iodide stain apoptotic cells?
How can I tell if my cell sample is dying?