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Z-DEVD-ProRed™ 620
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Application notes
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 DEVD-ProRed™ substrate is specific for detecting caspases 3 and 7.
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.
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
1. Z-DEVD-ProredTM 620 stock solution (10 mM):
Add 65 µL of DMSO into the vial of 1 mg Z-DEVD-ProredTM 620 to make 10 mM stock solution.
PREPARATION OF WORKING SOLUTION
Caspase 3/7 assay solution (2X):
Mix 50 µL Z-DEVD-ProRed™ 620 stock solution (10 mM), 100 µL DTT (1M), 400 µL EDTA (100 mM) and 10 mL Tris Buffer (20 mM), pH =7.4.
SAMPLE EXPERIMENTAL PROTOCOL
- Mix equal volume of the caspase 3/7 standards or samples with 2X caspase 3/7 assay reaction solution and incubate at room temperature for at least 1 hour.
- Monitor the fluorescence increase at Ex/Em = 535/620 nm.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of Z-DEVD-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 | 63.877 µL | 319.387 µL | 638.774 µL | 3.194 mL | 6.388 mL |
| 5 mM | 12.775 µL | 63.877 µL | 127.755 µL | 638.774 µL | 1.278 mL |
| 10 mM | 6.388 µL | 31.939 µL | 63.877 µL | 319.387 µL | 638.774 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
Product family
| Name | Excitation (nm) | Emission (nm) |
| Z-IETD-ProRed™ 620 | 532 | 619 |
| Z-LEHD-ProRed™ 620 | 532 | 619 |
Citations
View all 2 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 (2016): 181--192
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 (2016): 181--192
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
Fisetin induces apoptosis in human cervical cancer HeLa cells through ERK1/2-mediated activation of caspase-8-/caspase-3-dependent pathway
Authors: Ying TH, Yang SF, Tsai SJ, Hsieh SC, Huang YC, Bau DT, Hsieh YH.
Journal: Arch Toxicol (2012): 263
Authors: Ying TH, Yang SF, Tsai SJ, Hsieh SC, Huang YC, Bau DT, Hsieh YH.
Journal: Arch Toxicol (2012): 263
Andrographolide induces apoptosis in B16F-10 melanoma cells by inhibiting NF-kappaB-mediated bcl-2 activation and modulating p53-induced caspase-3 gene expression
Authors: Pratheeshkumar P, Sheeja K, Kuttan G.
Journal: Immunopharmacol Immunotoxicol (2012): 143
Authors: Pratheeshkumar P, Sheeja K, Kuttan G.
Journal: Immunopharmacol Immunotoxicol (2012): 143
ECRG4 is a negative regulator of caspase-8-mediated apoptosis in human T-leukemia cells
Authors: Matsuzaki J, Torigoe T, Hirohashi Y, Kamiguchi K, Tamura Y, Tsukahara T, Kubo T, Takahashi A, Nakazawa E, Saka E, Yasuda K, Takahashi S, Sato N.
Journal: Carcinogenesis (2012): 996
Authors: Matsuzaki J, Torigoe T, Hirohashi Y, Kamiguchi K, Tamura Y, Tsukahara T, Kubo T, Takahashi A, Nakazawa E, Saka E, Yasuda K, Takahashi S, Sato N.
Journal: Carcinogenesis (2012): 996
Proteasome inhibition can impair caspase-8 Activation upon sub-maximal Stimulation of apoptotic tumour necrosis factor-related apoptosis inducing ligand (TRAIL) signalling
Authors: Laussmann MA, Passante E, Hellwig CT, Tomiczek B, Flanagan L, Prehn JH, Huber HJ, Rehm M.
Journal: J Biol Chem. (2012)
Authors: Laussmann MA, Passante E, Hellwig CT, Tomiczek B, Flanagan L, Prehn JH, Huber HJ, Rehm M.
Journal: J Biol Chem. (2012)
Caspase-2 is an initiator caspase responsible for pore-forming toxin-mediated apoptosis
Authors: Imre G, Heering J, Takeda AN, Husmann M, Thiede B, Zu Heringdorf DM, Green DR, van der Goot FG, Sinha B, Dotsch V, Rajalingam K.
Journal: EMBO J. (2012)
Authors: Imre G, Heering J, Takeda AN, Husmann M, Thiede B, Zu Heringdorf DM, Green DR, van der Goot FG, Sinha B, Dotsch V, Rajalingam K.
Journal: EMBO J. (2012)