(Z-DEVD)2-R110

Since highly purified rhodamine 110 (R110)-derived substrates are locked in the lactone configuration they are colorless and non-fluorescent. Cleavage of R110 peptides by caspases generates strongly fluorescent R110 that can be monitored fluorimetrically at 510-530 nm with excitation of 488 nm, the most common excitation light source used in fluorescence instruments. R110-derived caspase substrates are probably the most sensitive indicators widely used for the fluorimetric detection of various caspase activities. This R110 substrate is specific for caspases 3 and 7. Our R110-based substrates are highly purified to eliminate the trace amount of free R110 that is not detectable by HPLC, but causes significant assay background.

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)₂-R110 stock solution (10 mM):
Add 65 µL of DMSO into the vial of 1 mg (Z-DEVD)₂-R110 to make 10 mM stock solution.

PREPARATION OF WORKING SOLUTION

Caspase 3/7 assay solution (2X):
Mix 50 µL (Z-DEVD)₂-R110 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 = 490/525 nm.

Spectrum

Open in Advanced Spectrum Viewer

Citations

View all 2 citations: Citation Explorer

Presented by: Martine Ballinger (B. Sc.) Born in: Westminster, London Oral examination: 4th October
Authors: Ballinger, Martine
Journal: (2021)

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

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