| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Aminoglycoside
Puromycin-d3 dihydrochloride, like its non-deuterated parent, targets the ribosome. Puromycin specifically inhibits peptidyl transferase activity in both prokaryotic and eukaryotic ribosomes by mimicking the 3'-end of aminoacyl-tRNA. It is incorporated into the growing polypeptide chain, causing premature chain termination and disrupting protein synthesis. The deuterium labeling does not alter the mechanism of action but serves as an internal standard for quantification. |
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers for quantification throughout the drug development process. Due to its potential to alter the pharmacokinetic and metabolic characteristics of medications, deuteration has drawn attention[1].
Puromycin dihydrochloride (the non-deuterated parent) is a potent inhibitor of protein synthesis in vitro. It is widely used as a selection agent for cells expressing the puromycin resistance gene (pac), which encodes puromycin N-acetyl-transferase. The deuterated version is not used for biological activity assessment; it is used as an analytical standard. Puromycin-d3 is the deuterium-labeled form used as an internal standard for quantitation. |
| ln Vivo |
In vivo, puromycin is used as a research tool to study protein synthesis. The non-deuterated parent, puromycin dihydrochloride, is an antibiotic that inhibits protein synthesis. It is not typically administered systemically due to toxicity but is used in cell culture as a selection agent. The deuterated version is used as an internal standard for PK studies of puromycin and its analogs.
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| Enzyme Assay |
Puromycin-d3 dihydrochloride 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 protein precipitation and analyzed on an LC-MS/MS system. Puromycin is detected using specific MRM transitions; the d3 internal standard has a mass shift of +3 Da. The peak area ratio is used for quantification.
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| Cell Assay |
Puromycin-d3 is not used in typical cell-based assays. The non-deuterated parent, puromycin, is used as a selection agent. HEK293 or HeLa cells expressing the puromycin resistance gene are cultured in DMEM + 10% FBS. Puromycin dihydrochloride is added to the medium at 0.5-10 microg/mL. Viable resistant cells survive, while non-resistant cells die within 2-4 days. Selection efficiency is confirmed by staining with crystal violet or by MTT assay.
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| Animal Protocol |
Puromycin-d3 is used as an internal standard for PK studies. Male Sprague-Dawley rats (200-250 g) are administered a single intravenous or intraperitoneal dose of puromycin dihydrochloride (1-10 mg/kg). Blood samples are collected at various time points. Plasma is separated and processed with a fixed concentration of the d3 internal standard. PK parameters are calculated using non-compartmental analysis. Puromycin-d3 itself is not administered in vivo.
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| ADME/Pharmacokinetics |
Puromycin-d3 dihydrochloride has a molecular weight of 547.45 (free base: 474.53) and the molecular formula C22H28D3Cl2N7O5. It is an off-white to light yellow solid powder with ≥98% purity. The powder should be stored at -20degC for up to 3 years. In solvent, it is stable for 6 months at -80degC. The compound should be protected from light and moisture.
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| Toxicity/Toxicokinetics |
No specific toxicity data for the deuterated version are available. The non-deuterated parent, puromycin, is toxic to both eukaryotic and prokaryotic cells due to its inhibition of protein synthesis. It causes cell death in rapidly dividing cells and is not suitable for in vivo therapeutic use. As an analytical standard, the deuterated version is used in trace amounts and does not pose additional safety risks.
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| References | |
| Additional Infomation |
Puromycin-d3 dihydrochloride (CL13900-d3 dihydrochloride) is a stable isotope-labeled research compound used as an internal standard for LC-MS/MS quantification of puromycin. The parent compound, puromycin, is an aminoglycoside antibiotic that inhibits protein synthesis. This product is for research use only and is not for human therapeutic applications.
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| Molecular Formula |
C22H28D3CL2N7O5
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|---|---|
| Molecular Weight |
547.45
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| Related CAS # |
Puromycin dihydrochloride;58-58-2
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| Appearance |
Off-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) |
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 | 1.8267 mL | 9.1333 mL | 18.2665 mL | |
| 5 mM | 0.3653 mL | 1.8267 mL | 3.6533 mL | |
| 10 mM | 0.1827 mL | 0.9133 mL | 1.8267 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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