| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
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| Targets |
L-Isoleucine targets branched-chain amino acid (BCAA) metabolic pathways. As an essential amino acid, it is incorporated into proteins and serves as a precursor for gluconeogenesis. Isoleucine activates the mTOR signaling pathway, regulating protein synthesis and cell growth. The compound is used to study BCAA metabolism and related metabolic disorders.
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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 cause for concern [1].
In vitro, L-isoleucine stimulates protein synthesis in muscle cells via mTOR activation. It promotes cell proliferation and differentiation. The deuterated form is used to trace isoleucine incorporation into proteins and other biomolecules in cell culture. Cellular uptake of labeled isoleucine is quantified by LC-MS/MS. The compound's effects on mTOR signaling are assessed by Western blot for phosphorylated S6K and 4E-BP1. |
| ln Vivo |
In vivo, L-isoleucine is essential for muscle protein synthesis, energy production, and glucose homeostasis. The deuterated form is used in metabolic tracing studies to investigate isoleucine metabolism in animal models. It is particularly useful in studying BCAA metabolism in conditions such as obesity, diabetes, and maple syrup urine disease. Isoleucine supplementation has been shown to improve glucose metabolism.
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| Enzyme Assay |
In vitro enzyme assays for branched-chain aminotransferase (BCAT) involve incubating the enzyme with L-Isoleucine-d10 and α-ketoglutarate in phosphate buffer at pH 7.4. The reaction product, α-keto-β-methylvalerate, is quantified by LC-MS/MS. Kinetic parameters (Km, Vmax) are determined by varying substrate concentrations. The deuterated substrate allows for precise quantification and tracing of the reaction products. The assay is typically performed at 37°C.
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| Cell Assay |
For in vitro cell assays, cells are cultured in isoleucine-free medium supplemented with L-Isoleucine-d10 at concentrations of 0.1-10 mM. Protein synthesis is assessed by measuring the incorporation of labeled isoleucine into newly synthesized proteins using LC-MS/MS. Cell proliferation is evaluated by MTT assay. mTOR signaling is analyzed by Western blot. Cellular uptake and metabolism of the labeled isoleucine are quantified by LC-MS/MS.
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| Animal Protocol |
For in vivo animal studies, L-Isoleucine-d10 is administered to rodents via oral gavage or intraperitoneal injection at doses of 50-200 mg/kg. Blood and tissue samples are collected at various time points for pharmacokinetic analysis. Plasma and tissue isoleucine levels are quantified by LC-MS/MS. Metabolic flux analysis traces the incorporation of deuterium into downstream metabolites and proteins. Stable isotope tracing is performed to study BCAA metabolism in various tissues.
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| ADME/Pharmacokinetics |
L-Isoleucine-d10 has a molecular weight of 141.23. It is soluble in water and physiological buffers. The compound is stable under recommended storage conditions (powder at -20°C). The deuterium labeling provides a mass shift of +10 Da for MS detection. It is absorbed via amino acid transporters and metabolized through the BCAA catabolic pathway. The compound is stable as a powder at -20°C for 3 years.
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| Toxicity/Toxicokinetics |
L-Isoleucine is generally recognized as safe at physiological concentrations. The deuterated form is not expected to exhibit additional toxicity. High doses may cause hyperaminoacidemia. The compound is for research use only, not for human therapeutic applications. No specific toxicity data are available for the deuterated analog; safety profiles are extrapolated from L-isoleucine, which is an essential amino acid.
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| References | |
| Additional Infomation |
L-Isoleucine-d10 (CAS# 35045-71-7) is a stable isotope-labeled compound used primarily as a tracer in metabolic studies and protein quantification. It has not been investigated in clinical trials nor approved as a therapeutic drug. The compound is widely used in research on BCAA metabolism, protein synthesis, and metabolic disorders such as maple syrup urine disease and diabetes. The deuterated form enables precise quantification of isoleucine in complex biological matrices.
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| Molecular Formula |
C6H3D10NO2
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|---|---|
| Molecular Weight |
141.23
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| Exact Mass |
141.157
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| CAS # |
35045-71-7
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| Related CAS # |
L-Isoleucine;73-32-5
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| PubChem CID |
101865217
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| Appearance |
White to off-white solid powder
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| LogP |
-1.7
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
9
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| Complexity |
103
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C(O)(=O)[C@]([2H])([C@@]([2H])(C[2H])C([2H])([2H])C([2H])([2H])[2H])N([2H])[2H]
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| InChi Key |
AGPKZVBTJJNPAG-VQWSQZATSA-N
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| InChi Code |
InChI=1S/C6H13NO2/c1-3-4(2)5(7)6(8)9/h4-5H,3,7H2,1-2H3,(H,8,9)/t4-,5-/m0/s1/i1D3,2D3,3D2,4D,5D
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| Chemical Name |
(2S,3S)-2-amino-2,3,4,4,5,5,5-heptadeuterio-3-(trideuteriomethyl)pentanoic acid
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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: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 | 7.0806 mL | 35.4032 mL | 70.8065 mL | |
| 5 mM | 1.4161 mL | 7.0806 mL | 14.1613 mL | |
| 10 mM | 0.7081 mL | 3.5403 mL | 7.0806 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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