Z-IETD-ProRed™ 620

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	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 SDS Protocol

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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Spectrum

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Spectral properties

Excitation (nm)	532
Emission (nm)	619

Product Family

Name	Excitation (nm)	Emission (nm)
Z-DEVD-ProRed™ 620	532	619
Z-LEHD-ProRed™ 620	532	619

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)

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

Dopaminochrome induces caspase-independent apoptosis in the mesencephalic cell line, MN9D
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

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

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

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

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

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

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)

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