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| 5mg |
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| 10mg |
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| Targets |
SAG-d3 targets the Smoothened (Smo) receptor, a key signal transducer in the Hedgehog (Hh) signaling pathway. The unlabeled parent compound, SAG, is a potent agonist of Smo, with an EC50 of 3 nM and a Kd of 59 nM for binding. Activation of Smo by SAG initiates a signaling cascade that leads to the activation of Gli transcription factors and subsequent expression of Hh target genes. SAG-d3 shares the same receptor target and is used as a tracer for analytical purposes.
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| ln Vitro |
SAG-d3 is not typically used to measure biological activity, as its unlabeled counterpart (SAG) is used for that purpose. SAG is a potent Smoothened agonist (EC50 = 3 nM) widely used to study the Hedgehog signaling pathway in developmental biology and cancer research. By activating Smo, SAG prevents the proteolytic processing of Gli proteins, allowing them to translocate to the nucleus and activate transcription. SAG-d3 serves as an internal standard for quantifying SAG in cells or tissues.
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| ln Vivo |
In vivo, SAG has been shown to activate the Hedgehog pathway, leading to effects on cell proliferation and differentiation. The deuterated version, SAG-d3, is not administered for therapeutic purposes but is used as an internal standard for pharmacokinetic (PK) studies. Accurate quantification of SAG levels in plasma and tissues is crucial for understanding its absorption, distribution, metabolism, and excretion (ADME) properties.
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| Enzyme Assay |
SAG-d3 is not used in an enzyme binding assay; it is an internal standard for LC-MS/MS. A fixed concentration (e.g., 10-50 ng/mL) of the d3 internal standard is added to biological samples (e.g., plasma, tissue homogenates). Samples are extracted by liquid-liquid extraction (e.g., with ethyl acetate) or protein precipitation. The extract is analyzed by LC-MS/MS with a C18 column and a mobile phase of water/acetonitrile (0.1% formic acid). Detection is performed in positive ion mode using specific MRM transitions. The d3 internal standard has a mass shift of +3 Da from the parent SAG. The peak area ratio is used for quantification.
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| Cell Assay |
In a typical cell-based assay, the unlabeled SAG is used to activate the Hedgehog pathway. For example, SHH-Light II cells (NIH3T3 fibroblasts with a Gli-luciferase reporter) are seeded in 96-well plates. Cells are treated with SAG (1-1000 nM) for 24-48 h. Luciferase activity is measured to determine the EC50. To measure cellular uptake, cells are treated with SAG, and the lysates are analyzed by LC-MS/MS using SAG-d3 as an internal standard. The concentration of SAG internalized by the cells can be quantified from the peak area ratio.
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| Animal Protocol |
SAG-d3 is used as an internal standard for PK studies of SAG in animal models. A standard protocol: female BALB/c mice (6-8 weeks) are administered a single oral (p.o., 5-20 mg/kg) or intravenous (i.v., 1-5 mg/kg) dose of SAG. Blood samples are collected at multiple time points (0-24 h). Plasma is separated and spiked with a fixed concentration of SAG-d3. After sample extraction (e.g., protein precipitation with acetonitrile), the samples are analyzed by LC-MS/MS. PK parameters (Cmax, Tmax, AUC, t½, CL, Vd, F%) are calculated using non-compartmental analysis.
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| ADME/Pharmacokinetics |
SAG-d3 has a molecular weight of 493.08 and a molecular formula of C28H25D3ClN3OS. The powder should be stored at -20degC for up to 3 years, protected from light and stored under nitrogen to prevent degradation. In solvent, it is stable for several months at -20degC. For in vivo use, the unlabeled SAG is typically formulated in DMSO or a vehicle containing PEG300 and Tween 80.
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| Toxicity/Toxicokinetics |
The unlabeled parent compound, SAG, is used for research purposes only and has no approved clinical indications. High concentrations of SAG can cause teratogenic effects by dysregulating Hedgehog signaling, which is critical for embryonic development. The deuterated version is used in trace amounts as an analytical standard and does not pose additional safety risks beyond standard chemical handling. Standard safety precautions for handling research chemicals should be followed.
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| References | |
| Additional Infomation |
SAG-d3 is a research-grade stable isotope-labeled compound and is not approved for clinical use. It is used as an internal standard for the quantification of SAG, a potent Smoothened (Smo) receptor agonist widely used in Hedgehog signaling research. This product is for research use only and not for human therapeutic applications. Store as a powder at -20degC, protected from light.
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| Molecular Formula |
C28H25D3CLN3OS
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| Molecular Weight |
493.08
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| Related CAS # |
SAG;912545-86-9
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| Appearance |
White to light yellow 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 :~100 mg/mL (~202.81 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 | 2.0281 mL | 10.1403 mL | 20.2807 mL | |
| 5 mM | 0.4056 mL | 2.0281 mL | 4.0561 mL | |
| 10 mM | 0.2028 mL | 1.0140 mL | 2.0281 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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