| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Targets |
Z-IETD-AFC targets caspase-8, an initiator caspase that plays a critical role in the extrinsic apoptosis pathway. Caspase-8 is activated upon the binding of death ligands to death receptors (e.g., Fas, TNFR1). Activated caspase-8 cleaves downstream effector caspases (caspase-3, -7), leading to apoptosis. By serving as a substrate for caspase-8, Z-IETD-AFC allows for the quantification of caspase-8 activity in cell lysates or in vitro. The compound is also cleaved by granzyme B, a serine protease involved in cytotoxic T lymphocyte-mediated apoptosis.
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| ln Vitro |
In vitro, Z-IETD-AFC is used as a fluorogenic substrate to measure caspase-8 activity. The compound is incubated with cell lysates or purified caspase-8 enzyme, and the release of AFC is measured fluorometrically at excitation/emission wavelengths of approximately 400/505 nm. The increase in fluorescence is proportional to caspase-8 activity. The substrate's activity is concentration-dependent, with effective concentrations typically in the micromolar range. Its specificity for caspase-8 over other caspases makes it a valuable tool for studying caspase-8-mediated apoptosis.
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| ln Vivo |
In vivo, Z-IETD-AFC is not used as a therapeutic agent but as a research tool for measuring caspase-8 activity in tissue samples. The compound is used in ex vivo studies to assess apoptosis in tissues from animal models of disease. Its in vivo applications are limited to research settings. The compound is for research use only and is not approved for human therapeutic applications.
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| Enzyme Assay |
The in vitro caspase-8 activity assay for Z-IETD-AFC typically uses cell lysates or purified caspase-8 enzyme. The assay is performed in 96-well plates with the substrate (typically 10-200 µM) and varying concentrations of the sample in assay buffer. The reaction is initiated by adding the substrate and incubated at 37°C for 1-2 hours. The release of AFC is measured fluorometrically at excitation/emission wavelengths of 400/505 nm. The fluorescence is proportional to caspase-8 activity. For inhibition studies, varying concentrations of a caspase-8 inhibitor are added to the reaction. Positive controls (e.g., recombinant caspase-8) and negative controls (buffer only, caspase-8 inhibitor) are included in each assay run.
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| Cell Assay |
For in vitro cellular assays, cells are treated with apoptotic stimuli to induce caspase-8 activation. Cell lysates are prepared, and caspase-8 activity is measured using Z-IETD-AFC as described above. Cell viability is assessed using MTT or CellTiter-Glo assays. For mechanism studies, the effects of caspase-8 inhibitors or modulators on caspase-8 activity are investigated. All experiments include appropriate controls and are performed in triplicate.
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| Animal Protocol |
For in vivo studies, Z-IETD-AFC is not typically administered to animals. It is used in ex vivo studies on tissue samples obtained from animals. Tissues are harvested, lysed, and caspase-8 activity is measured using the substrate. The compound may be used in models of apoptosis, neurodegeneration, or cancer. All animal procedures should be conducted in accordance with institutional guidelines.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of Z-IETD-AFC are not relevant, as it is a research reagent rather than a therapeutic agent. The compound has a molecular weight of 805.75 and is a peptide substrate. It is used in vitro and ex vivo for biochemical assays. The compound should be stored protected from light and moisture to prevent degradation.
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| Toxicity/Toxicokinetics |
The toxicology of Z-IETD-AFC is not relevant, as it is a research reagent used in vitro at low concentrations. The compound is not intended for human or animal consumption. Standard laboratory safety precautions should be followed when handling the compound. It is for research use only and is not approved for human therapeutic use.
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| References | |
| Additional Infomation |
Z-IETD-AFC is a fluorogenic substrate for caspase-8 and granzyme B. It contains the caspase-8 recognition sequence IETD and the fluorophore AFC. Upon cleavage, AFC is released, producing a fluorescent signal. It is used to measure caspase-8 activity in apoptosis research. The compound is not approved for human use and is intended for research purposes only. It is available as a high-purity research reagent for laboratory use.
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| Molecular Formula |
C37H42N5O13F3
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|---|---|
| Molecular Weight |
821.750280000001
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| Exact Mass |
821.273
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| CAS # |
219138-02-0
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| PubChem CID |
71312079
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| Appearance |
White to off-white solid powder
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| LogP |
3.902
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| Hydrogen Bond Donor Count |
8
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| Hydrogen Bond Acceptor Count |
16
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| Rotatable Bond Count |
20
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| Heavy Atom Count |
58
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| Complexity |
1550
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| Defined Atom Stereocenter Count |
6
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| SMILES |
CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(=O)O)C(=O)NC1=CC2=C(C=C1)C(=CC(=O)O2)C(F)(F)F)NC(=O)OCC3=CC=CC=C3
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| InChi Key |
RCIFCAJYIPUCJW-QXPVPWKFSA-N
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| InChi Code |
InChI=1S/C37H42F3N5O13/c1-4-18(2)30(45-36(56)57-17-20-8-6-5-7-9-20)34(54)42-24(12-13-27(47)48)32(52)44-31(19(3)46)35(55)43-25(16-28(49)50)33(53)41-21-10-11-22-23(37(38,39)40)15-29(51)58-26(22)14-21/h5-11,14-15,18-19,24-25,30-31,46H,4,12-13,16-17H2,1-3H3,(H,41,53)(H,42,54)(H,43,55)(H,44,52)(H,45,56)(H,47,48)(H,49,50)/t18-,19+,24-,25-,30-,31-/m0/s1
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| Chemical Name |
(4S)-5-[[(2S,3R)-1-[[(2S)-3-carboxy-1-oxo-1-[[2-oxo-4-(trifluoromethyl)chromen-7-yl]amino]propan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-4-[[(2S,3S)-3-methyl-2-(phenylmethoxycarbonylamino)pentanoyl]amino]-5-oxopentanoic acid
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O : ~5 mg/mL (~6.08 mM)
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.2169 mL | 6.0846 mL | 12.1692 mL | |
| 5 mM | 0.2434 mL | 1.2169 mL | 2.4338 mL | |
| 10 mM | 0.1217 mL | 0.6085 mL | 1.2169 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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