| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Pyridinium bisretinoid A2E-d4 TFA is a non-toxic, isotopically enriched variant of A2E that shares the same biological properties. The unlabeled parent compound, A2E, is an autofluorescent pigment that targets cellular membranes and mitochondria. Blue-light photoactivation of A2E mediates autophagy and the production of reactive oxygen species (ROS), which leads to lysosomal dysfunction and apoptosis in RPE cells.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as quantitative tracers while the drugs were being developed. Because deuteration may have an impact on a drug's pharmacokinetics and metabolic profile, it has drawn attention [1].
Pyridinium bisretinoid A2E is an initiator of blue-light-induced apoptosis. Photoactivation of A2E mediates autophagy and the production of reactive oxygen species (ROS). The compound is detected in lipofuscin granules of RPE cells and accumulates with age. A2E has been shown to activate the NLRP3 inflammasome and induce oxidative stress, contributing to RPE cell death and AMD pathogenesis. |
| ln Vivo |
The unlabeled parent compound, A2E, is involved in the pathogenesis of age-related macular degeneration (AMD) in vivo. A2E accumulation in RPE cells with age, combined with blue-light exposure, triggers photo-oxidative damage, inflammasome activation, and RPE apoptosis. The deuterated version is used as an analytical standard for quantifying A2E levels in biological samples from AMD models and patients.
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| Enzyme Assay |
A cell-free binding assay is not performed for A2E. However, a standard LC-MS/MS assay using A2E-d4 as an internal standard is used for quantification. A fixed concentration of A2E-d4 is added to biological samples (e.g., RPE lysates or plasma). Samples are extracted and injected onto an LC-MS/MS system. A2E is detected using a specific MRM transition; A2E-d4 serves as the internal standard. The peak area ratio is used for quantification.
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| Cell Assay |
Arpe-19 retinal pigment epithelial cells are seeded in 6-well plates and cultured in DMEM/F12 medium. Cells are treated with A2E (1-50 uM) for 16-24 hours, followed by blue-light exposure (430 nm, 2-4 J/cm2). After treatment, cell viability is measured by MTT assay. ROS production is quantified using DCFH-DA fluorescence. Apoptosis is assessed by Annexin V/PI staining or TUNEL assay. Autophagy markers such as LC3-II and p62 are analyzed by Western blot.
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| Animal Protocol |
Animal models of AMD are used to study A2E. Abca4-/- Rdh8-/- double knockout mice, which accumulate A2E in RPE cells, are exposed to blue light (430 nm, 8000 lux) for 30 minutes. A2E-d4 (1-10 mg/kg) is administered orally or intraperitoneally. RPE/choroid tissues are collected, and A2E levels are quantified by LC-MS/MS using A2E-d4 as an internal standard. RPE cell death and histological damage are assessed.
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| ADME/Pharmacokinetics |
Pyridinium bisretinoid A2E-d4 TFA has a molecular weight of 709.96 and the molecular formula C44H54D4F3NO3. It is a brown to reddish brown solid powder with ≥98% purity. It is soluble in DMSO at 4 mg/mL and should be stored at 4degC, protected from light, and stored under nitrogen. The deuterium labeling does not alter its physicochemical properties.
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| Toxicity/Toxicokinetics |
No detailed toxicity data are available for the deuterated form. The unlabeled A2E compound has been shown to induce RPE cell death via oxidative stress and inflammasome activation. A2E accumulation in RPE is associated with AMD pathology. As an analytical standard, the deuterated version is used in trace amounts and does not pose additional safety risks.
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| References |
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| Additional Infomation |
Pyridinium bisretinoid A2E-d4 TFA (A2E-d4 TFA) is a deuterium-labeled research standard used for the quantification of A2E, an initiator of blue-light-induced apoptosis involved in AMD. The TFA salt ensures stability and solubility for research use. This product is for research use only and is not for human therapeutic applications.
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| Molecular Formula |
C44H54D4F3NO3
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| Molecular Weight |
709.96
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| Related CAS # |
Pyridinium bisretinoid A2E TFA;1821308-73-9;Pyridinium bisretinoid A2E;173449-96-2
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| Appearance |
Brown to reddish brown solid powder
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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: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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) |
DMSO :~4 mg/mL (~5.63 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 | 1.4085 mL | 7.0427 mL | 14.0853 mL | |
| 5 mM | 0.2817 mL | 1.4085 mL | 2.8171 mL | |
| 10 mM | 0.1409 mL | 0.7043 mL | 1.4085 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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