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Z-IETD-pNA *CAS 219138-21-3*

Z-IETD-pNA is a colorimetric caspase-8/granzyme B substrate containing the benzyloxycarbonyl (Z) moiety. This substrate is hydrlyzed by caspase 8 to generate highly colored pNA that is measured at 405 nm by an absorption microplate reader or spectrophotometer.

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-IETD-pNA stock solution (10 mM):
Prepare a 10 mM stock solution in DMSO.

PREPARATION OF WORKING SOLUTION

2X caspase substrate (50 μM) assay solution as the following:

ComponentVolume
Z-IETD-pNA substrate stock solution50 µL
DTT (1M)100 µL
EDTA (100 mM)400 µL
Tris Buffer (20 mM), pH =7.410 mL
 
Total volume10.55 mL

SAMPLE EXPERIMENTAL PROTOCOL

  1. Mix equal volume of the caspase standards or samples with 2X caspase substrate assay solution.

  2. Incubate the solutions at room temperature for at least 1 hour.

  3. Monitor the fluorescence using a fluorescence microplate reader, or absorbance using an absorbance microplate reader.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Z-IETD-pNA *CAS 219138-21-3* 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 mM136.851 µL684.257 µL1.369 mL6.843 mL13.685 mL
5 mM27.37 µL136.851 µL273.703 µL1.369 mL2.737 mL
10 mM13.685 µL68.426 µL136.851 µL684.257 µL1.369 mL

Molarity calculator

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

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)
Z-IETD-AFC *CAS 219138-02-0*376482170001
Z-IETD-aminoluciferin36249918000

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 28, 2024

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Physical properties

Molecular weight

730.72

Solvent

DMSO

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

219138-21-3
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