| Size | Price | |
|---|---|---|
| 1mg | ||
| 5mg | ||
| 10mg | ||
| Other Sizes |
| Targets |
L-leucine activates the mechanistic target of rapamycin (mTOR) signaling pathway, serving as a critical nutrient sensor for protein synthesis and cell growth regulation.
|
|---|---|
| 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].
Leucine is used as a tracer in metabolic studies and for quantifying protein synthesis. The 13C-labeled form enables precise monitoring of carbon flux through pathways including the tricarboxylic acid (TCA) cycle and protein synthesis. |
| ln Vivo |
The 13C6-labeled leucine is used in stable-isotope tracing experiments to track metabolic fate of BCAAs. SILAV (Stable Isotope Labeling of Amino acids in Vivo) method yields information about protein metabolism in living organisms.
|
| Enzyme Assay |
Non-cell enzyme assays for branched-chain amino acid metabolism involve incubating purified BCAT1 or BCAT2 enzymes with Leucine-13C6 as substrate. The reaction mixture contains 50 mM Tris-HCl (pH 7.8), 0.5 mM alpha-ketoglutarate, 1 mM NAD+, and 5-50 uM Leucine-13C6. After incubation at 37degC for 10-30 minutes, the reaction is stopped by adding 10% trichloroacetic acid. Samples are centrifuged and the supernatant is derivatized with dansyl chloride or analyzed directly by LC-MS/MS to quantify the production of 13C-labeled alpha-ketoisocaproate and monitoring mass shifts corresponding to the 6-carbon labeling pattern.
|
| Cell Assay |
For metabolic flux analysis, cells are cultured in medium containing Leucine-13C6 as the sole leucine source at 0.4-0.8 mM concentration for 12-48 hours. At designated time points, cells are washed with cold PBS, harvested, and metabolites are extracted using 80% methanol. After evaporation, samples are reconstituted and analyzed by LC-MS/MS in full-scan mode to detect M+6 mass shifts. Data processing involves correction for natural abundance and calculation of isotopologue distributions. For protein synthesis studies, cells are pulsed with Leucine-13C6 for various times, and after lysis, 13C enrichment in proteins is measured by mass spectrometry following trypsin digestion.
|
| Animal Protocol |
In animal tracer studies, mice or rats receive an intravenous bolus or continuous infusion of Leucine-13C6 at doses typically ranging from 10-50 mg/kg. For SILAV protocols, animals receive a flooding dose of 13C6-leucine to equilibrate precursor pools. Blood samples are collected via tail vein at time points from 5 minutes to 24 hours. Plasma is separated and amino acids are derivatized. The enrichment of 13C-leucine is measured by gas chromatography-mass spectrometry (GC-MS) or LC-MS/MS. Tissues including liver, muscle, and brain are harvested, frozen in liquid nitrogen, and analyzed for 13C incorporation into free amino acids and tissue proteins. The 13C enrichment data is used to calculate protein synthesis rates using established kinetic models.
|
| ADME/Pharmacokinetics |
As a stable isotope tracer, Leucine-13C6 is typically used in minimal quantities for metabolic studies and does not undergo formal pharmacokinetic evaluation. The parent compound L-leucine is absorbed rapidly from the gastrointestinal tract via amino acid transporters (LAT1, SLC7A5). Plasma leucine concentrations range from 100-300 uM in humans. Leucine is metabolized primarily in skeletal muscle, liver, and adipose tissue through transamination to alpha-ketoisocaproate, followed by oxidative decarboxylation via the branched-chain alpha-keto acid dehydrogenase complex. The elimination half-life is approximately 1-2 hours. Leucine is not excreted unchanged in significant amounts.
|
| Toxicity/Toxicokinetics |
The 13C-labeled form is non-radioactive and considered safe for research use at tracer doses. Unlabeled L-leucine has an established safety profile as an essential amino acid and dietary supplement. The acute oral LD50 in rodents is >5,000 mg/kg. At high doses exceeding physiological requirements, leucine may cause ammonia accumulation, hyperammonemia, and disruption of other amino acid levels. The 13C isotope is stable and non-toxic, with no additional hazard beyond the parent compound.
|
| References |
[1]. Russak EM, et al. Impact of Deuterium Substitution on the Pharmacokinetics of Pharmaceuticals. Ann Pharmacother. 2019;53(2):211-216.
[2]. Baoshan Xu, et al. Stimulation of mTORC1 with L-leucine rescues defects associated with Roberts syndrome. PLoS Genet. 2013;9(10):e1003857. [3]. Bruckbauer A, et al. Synergistic effects of leucine and resveratrol on insulin sensitivity and fat metabolism in adipocytes and mice. Nutr Metab (Lond). 2012 Aug 22;9(1):77. [4]. Rachdi L, et al. L-leucine alters pancreatic β-cell differentiation and function via the mTor signaling pathway. Diabetes. 2012 Feb;61(2):409-17. |
| Additional Infomation |
Leucine-13C6 is widely used as a stable isotopic tracer for metabolic flux analysis, protein turnover studies, muscle protein synthesis research, and BCAA metabolism investigations. It serves as a non-radioactive internal standard in mass spectrometry-based quantification and is employed in SILAC (Stable Isotope Labeling by Amino acids in Cell culture) proteomics and clinical tracer studies. The compound is also used to study mTOR signaling pathway activation and has applications in diabetes, obesity, and cachexia research.
|
| Molecular Formula |
13C6H13NO2
|
|---|---|
| Related CAS # |
L-Leucine;61-90-5
|
| Appearance |
Typically exists as solid at room temperature
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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.) |
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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
