| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Fat mass and obesity-associated protein (FTO).
|
|---|---|
| ln Vitro |
N6,2′-O-Dimethyladenosine is a reversible modification found at the +1 position adjacent to the 5′ mRNA cap, influencing cap-dependent translation. As a substrate, it undergoes FTO-catalyzed demethylation, which removes methyl groups from the RNA, impacting mRNA stability, translation efficiency, and protein expression. It is also a substrate for ALKBH5.
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| Enzyme Assay |
The FTO enzymatic assay is used to study demethylation. Recombinant FTO protein is incubated with N6,2′-O-Dimethyladenosine in a reaction buffer containing cofactors (alpha-ketoglutarate, Fe2+, ascorbate). The reaction products are analyzed by LC-MS/MS to quantify the formation of demethylated adenosine derivatives. This setup allows measurement of FTO catalytic activity and screening of FTO inhibitors.
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| Cell Assay |
To study its role in cells, N6,2′-O-Dimethyladenosine or its modified RNA transcripts are transfected into cells expressing FTO. Changes in the stability and translational efficiency of the modified RNA are monitored using qRT-PCR and reporter assays (e.g., luciferase). Alternatively, global m6Am levels in mRNA are measured by LC-MS after treatment with FTO inhibitors.
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| Animal Protocol |
In vivo studies are conducted using genetically modified mouse models (e.g., FTO knockout or overexpressing mice). The levels of N6,2′-O-Dimethyladenosine in various tissues (especially brain and adipose) are quantified by LC-MS. Correlations between levels of this modification, body weight, and metabolic parameters are studied to understand the physiological role of FTO.
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| ADME/Pharmacokinetics |
As an endogenous modified nucleoside, N6,2′-O-Dimethyladenosine is metabolically unstable and is likely degraded by nucleotidases and phosphorylases. Its intracellular levels are tightly regulated by the balance between its incorporation into RNA via the transcription machinery and its demethylation by FTO and other eraser proteins.
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| Toxicity/Toxicokinetics |
No toxicity data are available as this is an endogenous RNA modification, not an exogenous drug. It is present in normal cells at low levels and is part of normal RNA metabolism. High levels of this modification may disrupt normal translation and are potentially hazardous, but no studies have established acute toxicity.
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| References | |
| Additional Infomation |
N(6),2'-O-dimethyladenosine is a methyladenosine compound in which two methyl groups are attached to the N(6) position of the adenine nucleobase and the O(2) position of the sugar, respectively. It is functionally related to adenosine.
This compound is a research chemical, not an approved drug. It has a key biological function as a regulator of gene expression, specifically in the context of the mRNA cap. By regulating cap-dependent translation, it influences various cellular processes including cell growth and differentiation. It is used as a standard in LC-MS assays for studying FTO activity and for quantifying m6Am levels in biological samples. |
| Molecular Formula |
C12H17N5O4
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|---|---|
| Molecular Weight |
295.29
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| Exact Mass |
295.128
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| CAS # |
57817-83-1
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| PubChem CID |
6453528
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| Appearance |
White to off-white solid powder
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| LogP |
0.2
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| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
21
|
| Complexity |
363
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
CNC1=C2C(=NC=N1)N(C=N2)[C@H]3[C@@H]([C@@H]([C@H](O3)CO)O)OC
|
| InChi Key |
GRYSXUXXBDSYRT-WOUKDFQISA-N
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| InChi Code |
InChI=1S/C12H17N5O4/c1-13-10-7-11(15-4-14-10)17(5-16-7)12-9(20-2)8(19)6(3-18)21-12/h4-6,8-9,12,18-19H,3H2,1-2H3,(H,13,14,15)/t6-,8-,9-,12-/m1/s1
|
| Chemical Name |
(2R,3R,4R,5R)-2-(hydroxymethyl)-4-methoxy-5-[6-(methylamino)purin-9-yl]oxolan-3-ol
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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: 250 mg/mL (846.63 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.47 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), Clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (8.47 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), Clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 900 μL of 20% SBE-β-CD saline and mix well. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.5 mg/mL (8.47 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), Clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3865 mL | 16.9325 mL | 33.8650 mL | |
| 5 mM | 0.6773 mL | 3.3865 mL | 6.7730 mL | |
| 10 mM | 0.3387 mL | 1.6933 mL | 3.3865 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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