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

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Chemical structure for Z-IETD-pNA *CAS 219138-21-3*
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5 mg 13413 $195

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Ex/Em (nm)408/None
CAS #219138-21-3
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Category Enzyme Detection
Peptidases and Proteases
Related Cell Apoptosis
Apoptosis and Cytotoxicity
Secondary Reagents
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.

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.

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Table 2. Enter any two values (mass, volume, concentration) to calculate the third.

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This protocol only provides a guideline, and should be modified according to your specific needs.

1.       General Solution Caspase Assays Using AMC, AFC, pNA, R110 and ProRed Substrates

1.1.     Prepare a 10 mM stock solution in DMSO.

1.2.     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

1.3.     Mix equal volume of the caspase standards or samples with 2X caspase substrate assay solution (from Step 1.2), and incubate the solutions at room temperature for at least 1 hour.

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

2.       Cell Caspase Assays Using Cell-Permeable FMK Caspase Probes

2.1.     Prepare a 2-5 mM stock solution in DMSO.

2.2.     Treat cells as desired.

2.3.     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).

2.4.     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.

2.5.     Wash the cells with HHBS for at least once.

2.6.     Monitor the fluorescence intensity by a flow cytometer, a fluorescence microscope or a fluorescence microplate reader.

3.       Cell Caspase Assays Using Cell-Permeable FMK Caspase Probes (For #13470-13476 only)

3.1.     Prepare a 250X stock solution by adding 50 µL DMSO into the vial.

3.2.     Treat cells as desired.

3.3.     Add 250 X DMSO stock solution (from Step 3.1) into the cell solution at a 1:250 ratio (such as 2 uL to 500 uL cells), and incubate the cells in a 37°C, 5% CO2 incubator for 1 hour.

3.4.     Wash the cells with HHBS for at least once.

3.5.     Monitor the fluorescence intensity by flow cytometer, fluorescence microscopy or fluorescent microplate reader.

References & Citations

pH-Assisted surface functionalization of selenium nanoparticles with curcumin to achieve enhanced cancer chemopreventive activity
Authors: Shaoxuan Yu, Yanru Wang, Wentao Zhang, Yuhuan Zhang, Wenxin Zhu, Yingnan Liu, Daohong Zhang, Jianlong Wang
Journal: RSC Advances (2016): 72213--72223

Additional Documents

Safety Data Sheet (SDS)

1. Enzyme Probes & Assay Kits
2. Cell Apoptosis & Proliferation

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