| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
The primary biological target is EBI2 (GPR183), an orphan GPCR expressed on B cells, T cells, dendritic cells, and other immune cells. 7α,25-OHC binds EBI2 with high affinity (Kd 450 pM) and activates Gαi-mediated signaling, directing immune cell migration and positioning within lymphoid follicles. The deuterated standard retains EBI2 agonist activity and serves as an analytical surrogate for quantification.
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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 effect on a drug's pharmacokinetics and metabolic properties, it is a matter for concern [3].
7α,25-OHC stimulates the migration of EBI2-expressing mouse B and T cells with half-maximum effective concentration values around 500 pM in vitro, while having no effect on EBI2-deficient cells. The d6-labeled standard shows comparable activity and is used as an isotope-dilution internal standard for LC-MS/MS quantification of 7α,25-OHC in biological matrices. |
| ln Vivo |
In vivo, B cells lacking EBI2 or normal B cells rendered insensitive through pre-treatment with 7α,25-OHC (1 μM; 1.5 h) display decreased migration to spleen follicular regions. The d6 standard supports in vivo quantification of 7α,25-OHC in plasma and lymphoid tissues and enables studies of oxysterol-mediated immune cell trafficking.
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| Enzyme Assay |
EBI2 radioligand binding assay: incubate membranes from EBI2-expressing cells with a radiolabeled EBI2 ligand in the presence of serially diluted 7α,25-OHC-d6; separate bound from free ligand by filtration and determine Kd or IC50. The d6 standard is quantified by LC-MS/MS to verify assay concentrations.
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| Cell Assay |
Cell-based EBI2 migration assay: culture EBI2-expressing B cells or transiently transfected cells, treat with serially diluted 7α,25-OHC-d6, and measure chemotaxis in a Transwell chamber; quantify migrated cells by flow cytometry. EBI2-dependent signaling is confirmed by comparing responses in EBI2-deficient cells.
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| Animal Protocol |
In vivo immune cell trafficking study: administer 7α,25-OHC-d6 or unlabeled 7α,25-OHC to mice, assess B cell and dendritic cell positioning in spleen and lymph nodes by immunohistochemistry or flow cytometry, and quantify 7α,25-OHC levels in tissues by LC-MS/MS.
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| References |
| Molecular Formula |
C27H40D6O3
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|---|---|
| Molecular Weight |
424.69
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| CAS # |
2260669-11-0
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| Related CAS # |
7α,25-Dihydroxycholesterol;64907-22-8
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| Appearance |
Typically exists as solid at room temperature
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| SMILES |
C[C@@]12[C@](CC[C@]2([H])[C@H](C)CCCC(C([2H])([2H])[2H])(O)C([2H])([2H])[2H])([H])[C@@]3([H])[C@@](CC1)([H])[C@@]4(C(C[C@H](CC4)O)=C[C@H]3O)C
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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 | 2.3547 mL | 11.7733 mL | 23.5466 mL | |
| 5 mM | 0.4709 mL | 2.3547 mL | 4.7093 mL | |
| 10 mM | 0.2355 mL | 1.1773 mL | 2.3547 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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