| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
A2/J2-isoprostanes contain a reactive alpha,beta-unsaturated carbonyl group that can undergo Michael addition reactions with thiol-containing biomolecules like glutathione (GSH) and proteins. This covalent modification of proteins is a key mechanism by which 15-A2t-Isoprostane exerts its biological effects. It can also signal through the thromboxane A2 (TP) receptor, inhibiting angiogenesis.
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| ln Vitro |
15-A2t-Isoprostane (10, 30 μM, 3 h) enhanced glutamate-induced cytotoxicity and reduced glutathione (GSH) levels in HT22 hippocampal cells[2]. 15-A2t-Isoprostane (10 μM, 24 h) inhibited VEGF-induced migration and tubule formation in human coronary artery endothelial cells[3].
15-A2t-Isoprostane (10, 30 microM, 3 h) enhances glutamate-induced cytotoxicity and reduces glutathione (GSH) levels in HT22 hippocampal cells. It also (10 microM, 24 h) inhibits VEGF-induced migration and tube formation in human coronary artery endothelial cells via activation of the TP receptor. This demonstrates its dual role as a cytotoxic and anti-angiogenic agent. |
| ln Vivo |
Following intravenous administration in rats, 80% of the radioactive dose from [3H]15-A2t-Isoprostane is excreted in urine as a polar conjugate. The major metabolite identified was a mercapturic acid sulfoxide, formed after conjugation with glutathione. Furthermore, it is formed endogenously in rats exposed to an oxidant stress, and the urinary metabolite can be used as a specific biomarker.
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| Enzyme Assay |
Not applicable. As a highly reactive compound that reacts directly with biomolecules, its effects are studied in cell-based assays. A standard protocol for detection of its urinary metabolite: Urine is extracted with organic solvents; however, the conjugate remains in the aqueous phase. The mercapturic acid sulfoxide metabolite is identified using high-pressure liquid chromatography coupled with mass spectrometry (HPLC/MS).
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| Cell Assay |
Cytotoxicity assay in HT22 mouse hippocampal cells: HT22 cells are treated with 15-A2t-Isoprostane (e.g., 10-30 microM) for 3 hours in the presence or absence of glutamate. Cell viability is then measured using an MTT assay. Reduced glutathione (GSH) levels in the cells can be measured using a fluorometric or colorimetric assay kit. This protocol allows for the assessment of the compound's neurotoxic potential.
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| Animal Protocol |
Urinary metabolite identification study in rats: To study the in vivo fate of 15-A2t-Isoprostane, a solution containing the compound spiked with a small amount of [3H]15-A2t-Isoprostane is administered intravenously to rats. Urine is then collected over a 24-hour period. The urine is extracted with organic solvents, and the remaining radioactivity in the aqueous phase is measured. The major metabolite is then identified by HPLC/MS.
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| ADME/Pharmacokinetics |
Following intravenous administration of 15-A2t-Isoprostane, 80% of the radioactivity is rapidly excreted in urine as a polar metabolite. The major urinary metabolite was identified as the mercapturic acid sulfoxide conjugate. This indicates that the compound has a very short half-life in circulation and is rapidly cleared from the body through conjugation with glutathione.
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| Toxicity/Toxicokinetics |
15-A2t-Isoprostane is a reactive electrophile that can form covalent adducts with proteins, a process that can lead to cellular dysfunction. Its administration to animals is used to understand its potential role as a downstream mediator of oxidative stress in diseases. It has been shown to be efficiently metabolized by HepG2 cells via conjugation with glutathione.
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| References |
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| Additional Infomation |
8-Isoprostaglandin A2 is a prostaglandin. Cyclopentenone isoprostaglandin
15-A2t-Isoprostane (CAS: 474391-66-7) is also known as 8-iso Prostaglandin A2. Its molecular formula is C20H30O4, with a molecular weight of 334.45. This isoprostane is a non-enzymatic, free radical-catalyzed product of arachidonic acid and serves as a biomarker of oxidative stress. |
| Molecular Formula |
C20H30O4
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|---|---|
| Molecular Weight |
334.45
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| Exact Mass |
334.214
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| CAS # |
474391-66-7
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| PubChem CID |
35020890
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| Appearance |
Typically exists as solids at room temperature
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| Density |
1.1±0.1 g/cm3
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| Boiling Point |
515.3±50.0 °C at 760 mmHg
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| Flash Point |
279.6±26.6 °C
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| Vapour Pressure |
0.0±3.0 mmHg at 25°C
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| Index of Refraction |
1.555
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| LogP |
3.25
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
12
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| Heavy Atom Count |
24
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| Complexity |
476
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| Defined Atom Stereocenter Count |
3
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| SMILES |
CCCCC[C@@H](/C=C/[C@H]1C=CC(=O)[C@H]1C/C=C\CCCC(=O)O)O
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| InChi Key |
MYHXHCUNDDAEOZ-UKUWKSPLSA-N
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| InChi Code |
InChI=1S/C20H30O4/c1-2-3-6-9-17(21)14-12-16-13-15-19(22)18(16)10-7-4-5-8-11-20(23)24/h4,7,12-18,21H,2-3,5-6,8-11H2,1H3,(H,23,24)/b7-4-,14-12+/t16-,17-,18-/m0/s1
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| Chemical Name |
(Z)-7-[(1S,2S)-2-[(E,3S)-3-hydroxyoct-1-enyl]-5-oxocyclopent-3-en-1-yl]hept-5-enoic acid
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| Synonyms |
8-iso Prostaglandin A2
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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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 2.9900 mL | 14.9499 mL | 29.8998 mL | |
| 5 mM | 0.5980 mL | 2.9900 mL | 5.9800 mL | |
| 10 mM | 0.2990 mL | 1.4950 mL | 2.9900 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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