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

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

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

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

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)

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)

Inhibitory effects of Scutellaria baicalensis extract on hepatic stellate cells through inducing G2/M cell cycle arrest and activating ERK-dependent apoptosis via Bax and caspase pathway
Authors: Pan TL, Wang PW, Leu YL, Wu TH, Wu TS.
Journal: J Ethnopharmacol (2012): 829

Berberine inhibits norepinephrine-induced apoptosis in neonatal rat cardiomyocytes via inhibiting ROS-TNF-alpha-caspase signaling pathway
Authors: Lv XX, Yu XH, Wang HD, Yan YX, Wang YP, Lu DX, Qi RB, Hu CF, Li HM.
Journal: Chin J Integr Med. (2012)

Imaging apoptosis with positron emission tomography: 'bench to bedside' development of the caspase-3/7 specific radiotracer [(18)F]ICMT-11
Authors: Nguyen QD, Challapalli A, Smith G, Fortt R, Aboagye EO.
Journal: Eur J Cancer (2012): 432

Musca domestica pupae lectin induces apoptosis in HepG2 cells through a NF-kappaB/p65-mediated caspase pathway
Authors: Nie J, Cao X, Zhou M, Zhang X, Zhang R, Niu L, Xia Y, Hou L, Wang C.
Journal: Bull Cancer (2012): 49

Caspase cleavage of viral proteins, another way for viruses to make the best of apoptosis
Authors: Richard A, Tulasne D.
Journal: Cell Death Dis (2012): e277