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Z-IETD-FMK, Caspase-8 Inhibitor
Z-IETD-FMK, Caspase-8 Inhibitor

Flow cytometric analysis of Jurkat cells (Human T-cell leukemia; ATCC TIB-152). Jurkat cells were preincubated with the following: no inhibitor (upper left and bottom left panels), 20 µM Z-IETD-FMK (upper center and bottom center panels) or 20 µM of a negative control inhibitor Z-FA-FMK (upper right and bottom right panels) for 30 minutes, and then either left untreated (bottom row) or treated with 4 µM of campthothecin for 3 hours (top row). Following incubation, cells were collected and stained with PE Annexin V (Cat. No. 559763) to identify cells undergoing apoptosis. The results indicate that in camthothecin treated cells, approximately 42% of the cells were induced to undergo apoptosis and the use of the caspase-8 inhibitor Z-IETD-FMK reduced the level of apoptosis to that observed in untreated controls. Cells treated with Z-FA-FMK (Cat. No. 550411) showed similar results to the treated cells without inhibitor, indicating that the control inhibitor did not attentuate apoptosis.

Flow cytometric analysis of Jurkat cells (Human T-cell leukemia; ATCC TIB-152). Jurkat cells were preincubated with the following: no inhibitor (upper left and bottom left panels), 20 µM Z-IETD-FMK (upper center and bottom center panels) or 20 µM of a negative control inhibitor Z-FA-FMK (upper right and bottom right panels) for 30 minutes, and then either left untreated (bottom row) or treated with 4 µM of campthothecin for 3 hours (top row). Following incubation, cells were collected and stained with PE Annexin V (Cat. No. 559763) to identify cells undergoing apoptosis. The results indicate that in camthothecin treated cells, approximately 42% of the cells were induced to undergo apoptosis and the use of the caspase-8 inhibitor Z-IETD-FMK reduced the level of apoptosis to that observed in untreated controls. Cells treated with Z-FA-FMK (Cat. No. 550411) showed similar results to the treated cells without inhibitor, indicating that the control inhibitor did not attentuate apoptosis.

Product Details
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BD Pharmingen™
Fluorescence quantitation (Routinely Tested)
AB_2868882
Lyophilized in dimethyl sulfoxide (DMSO).
RUO


Preparation And Storage

Avoid multiple freeze-thaws of product.

Store the lyophilized Z-IETD-FMK inhibitor at -20°C. Reconstitute the Z-IETD-FMK inhibitor in DMSO before use. The reconstituted Z-IETD-FMK inhibitor may be stored in small aliquots at -20°C.

Recommended Assay Procedures

The Z-IETD-FMK inhibitor is designed to be used in both in vivo and in vitro cell based assays to measure the inhibition of apoptosis. Reconstitute 1.0 mg of the Z-IETD-FMK inhibitor in DMSO. A 10 mM stock solution may be made by dissolving 1.0 mg of Z-IETD-FMK in 150 µl DMSO. The final concentration of inhibitor may vary between experimental systems and investigators are encouraged to titrate the inhibitor for optimal performance. As a precautionary note, do not exceed a final DMSO concentration of 0.2% as higher levels may cause cellular toxicity and mask the effects of the caspase inhibitor.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
550380 Rev. 2
Antibody Details
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Members of the caspase family play key roles in inflammation and mammalian apoptosis. Z-IETD-FMK is an irreversible and cell permeable inhibitor of caspase-8. The peptide is O-methylated in the P1 position on aspartic acid providing enhanced stability and increased cell permeability. This inhibitor can be used to inhibit caspase-8 activity and to study events downstream of caspase-8 activation. Z-IETD-FMK has a molecular weight of 654 Daltons.

550380 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
550380 Rev.2
Citations & References
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Development References (3)

  1. Gregoli PA, Bondurant MC. Function of caspases in regulating apoptosis caused by erythropoietin deprivation in erythroid progenitors. J Cell Physiol. 1999; 178(2):133-143. (Biology). View Reference
  2. Thornberry NA, Lazebnik Y. Caspases: enemies within. Science. 1998; 281(5381):1312-1316. (Biology). View Reference
  3. Wang J, Zhen L, Klug MG, Wood D, Wu X, Mizrahi J. Involvement of caspase 3- and 8-like proteases in ceramide-induced apoptosis of cardiomyocytes. J Card Fail. 2000; 6(3):243-249. (Biology). View Reference
550380 Rev. 2

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

Non-IVD products are For Research Use Only. Not for use in diagnostic or therapeutic procedures.

 

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