| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
MitoTEMPOL primarily targets mitochondria, specifically complex I and complex III of the electron transport chain where superoxide is generated. The TPP+ moiety drives its accumulation into the mitochondrial matrix (50-100 fold higher than in cytosol). Once inside, the TEMPOL nitroxide group acts as a superoxide dismutase mimetic, catalyzing the dismutation of superoxide anions into hydrogen peroxide and molecular oxygen. Unlike its parent compound TEMPOL, MitoTEMPOL does not significantly affect cytosolic ROS levels at selective concentrations, allowing for targeted investigation of mitochondrial oxidative stress without confounding effects on cellular redox balance .
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| ln Vitro |
In cell-free assays, MitoTEMPOL demonstrates SOD mimetic activity comparable to native SOD enzyme. The ability to scavenge superoxide is measured using the xanthine/xanthine oxidase system coupled with cytochrome c reduction. MitoTEMPOL (0.1-100 uM) inhibits cytochrome c reduction in a dose-dependent manner, with an IC50 of approximately 10-30 uM. Electron paramagnetic resonance (EPR) spectroscopy confirms the nitroxide radical's stability and its redox cycling capability. Unlike non-targeted antioxidants, MitoTEMPOL shows minimal activity in cytosolic fractions when tested in cell lysates due to its high positive charge and selective mitochondrial accumulation .
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| ln Vivo |
In cell-based studies, MitoTEMPOL effectively reduces mitochondrial superoxide levels without affecting cytosolic ROS. HeLa cells or primary neurons are loaded with MitoSOX Red (a mitochondrial superoxide indicator) and treated with MitoTEMPOL (1-100 uM) for 30-60 minutes. Following treatment, cells are exposed to oxidative stress inducers such as rotenone (complex I inhibitor, 1 uM) or antimycin A (complex III inhibitor, 10 uM). MitoTEMPOL pretreatment reduces MitoSOX fluorescence by 50-80% in a dose-dependent manner, indicating effective superoxide scavenging. The compound also reduces mitochondrial DNA damage and preserves mitochondrial membrane potential (deltaΨm) measured by TMRM (tetramethylrhodamine methyl ester) staining. Cytotoxicity (MTT assay) is not observed at concentrations below 100 uM .
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| Enzyme Assay |
A non-cell SOD mimetic activity assay uses the xanthine/xanthine oxidase system to generate superoxide radicals. A reaction mixture containing 50 uM xanthine, 50 uM cytochrome c (oxidized form, as a superoxide indicator), and varying concentrations of MitoTEMPOL (0.1-100 uM) is prepared in 50 mM potassium phosphate buffer (pH 7.8) containing 0.1 mM EDTA. The reaction is initiated by adding 0.01 U/mL xanthine oxidase. Reduction of cytochrome c (absorbance increase at 550 nm) is monitored kinetically for 2-3 minutes using a spectrophotometer. The superoxide scavenging activity is calculated as the percentage inhibition of cytochrome c reduction compared to control (no inhibitor). A standard curve using native SOD provides validation .
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| Cell Assay |
For mitochondrial superoxide measurement, cells are seeded in black-walled, clear-bottom 96-well plates (2×10⁴ cells/well) in appropriate culture medium and incubated overnight at 37degC with 5% CO2. Cells are pretreated with MitoTEMPOL (1-100 uM) for 30-60 minutes, then loaded with MitoSOX Red (5 uM in HBSS) for 10 minutes at 37degC in the dark. After washing with warm HBSS, cells are treated with an oxidative stress inducer (e.g., rotenone 1 uM, or antimycin A 10 uM) for 15-30 minutes. Fluorescence is measured using a plate reader (excitation 510 nm, emission 580 nm) or visualized by confocal microscopy. For quantification, fluorescence intensity is normalized to cell number or protein concentration .
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| Animal Protocol |
A mouse model of neuroinflammation or ischemia-reperfusion injury is commonly used. Adult male C57BL/6 mice (8-10 weeks, 20-25g) are subjected to transient middle cerebral artery occlusion (MCAO) for 60 minutes, followed by 24 hours of reperfusion. MitoTEMPOL (5-20 mg/kg) is administered intraperitoneally immediately before reperfusion, or intravenously (1-5 mg/kg) via tail vein injection. Following reperfusion, mice are euthanized, and brain tissues are collected. Infarct volume is measured by TTC (2,3,5-triphenyltetrazolium chloride) staining. Oxidative stress markers (malondialdehyde, 8-hydroxy-2'-deoxyguanosine, 4-HNE) and superoxide levels (dihydroethidium staining) are assessed in tissue homogenates .
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| ADME/Pharmacokinetics |
Pharmacokinetic studies in rodents show that MitoTEMPOL has moderate bioavailability. The TPP+ cation facilitates rapid cellular uptake and mitochondrial accumulation. Following intravenous administration (2-5 mg/kg) in mice, MitoTEMPOL reaches peak mitochondrial concentrations within 5-15 minutes. The compound has a short plasma half-life (t1/2 ~15-30 minutes) due to rapid tissue distribution and reduction of the nitroxide radical to the corresponding hydroxylamine. The hydroxylamine can be re-oxidized to the active nitroxide by cellular oxidants. The compound is cleared primarily via urinary excretion. For research use, MitoTEMPOL is stored as a powder at -20degC and is soluble in DMSO and water (10 mg/mL each) .
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| Toxicity/Toxicokinetics |
Toxicology data for MitoTEMPOL is limited. As a nitroxide radical, high concentrations may generate oxidative stress under certain conditions due to redox cycling. However, the selectivity of MitoTEMPOL for mitochondria generally minimizes off-target effects. In rodent studies, intraperitoneal administration of MitoTEMPOL at doses up to 50 mg/kg is well tolerated without significant behavioral or physiological abnormalities. The LD50 is estimated to be >200 mg/kg in mice. The TPP+ moiety at very high doses (not achieved with MitoTEMPOL) can inhibit mitochondrial function. Standard safety precautions apply: avoid inhalation, skin contact, and ingestion. The compound should be handled with gloves and lab coat .
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| References | |
| Additional Infomation |
MitoTEMPOL is a research tool, not an FDA-approved drug. It is one of several mitochondria-targeted antioxidants developed to study the role of mitochondrial superoxide in disease pathogenesis. MitoTEMPOL is often compared to MitoTEMPO (another mitochondria-targeted SOD mimetic with a different linker). The presence of a bromine atom in MitoTEMPOL allows for its detection by mass spectrometry or X-ray fluorescence. The compound is stable for up to 3 years when stored as a powder at -20degC and for 6 months when stored in solution at -80degC. The CAS number is 1101113-39-6, and purity is typically >98% by HPLC .
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| Molecular Formula |
C32H42BRNO2P*
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| Molecular Weight |
583.56
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| Exact Mass |
582.214
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| CAS # |
1101113-39-6
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| PubChem CID |
132285183
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| Appearance |
Brown to orange solid powder
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| LogP |
0
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
37
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| Complexity |
583
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C([P+](C1C=CC=CC=1)(C1C=CC=CC=1)C1C=CC=CC=1)CCCCOC1CC(C)(C)N([O])C(C)(C)C1.[Br-] |^1:31|
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| InChi Key |
NCMZESLTQCMFES-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C32H42NO2P.BrH/c1-31(2)25-27(26-32(3,4)33(31)34)35-23-15-8-16-24-36(28-17-9-5-10-18-28,29-19-11-6-12-20-29)30-21-13-7-14-22-30;/h5-7,9-14,17-22,27H,8,15-16,23-26H2,1-4H3;1H/q+1;/p-1
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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) |
DMF : ~50 mg/mL (~85.68 mM; with heating and sonication)
DMSO : ~50 mg/mL (~85.68 mM; with heating and sonication) Ethanol : ~50 mg/mL (~85.68 mM; with heating and sonication) |
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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 | 1.7136 mL | 8.5681 mL | 17.1362 mL | |
| 5 mM | 0.3427 mL | 1.7136 mL | 3.4272 mL | |
| 10 mM | 0.1714 mL | 0.8568 mL | 1.7136 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.
Link: https://clinicaltrials.gov/ct2/show/NCT06466655
Conditions:Middle-aged Adults|Major Depressive DisorderProducts are for research use only; We do not sell to patients
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