| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
Superoxide anion (O2•−). HKSOX-1r is a fluorescent sensor designed to specifically react with superoxide, a key reactive oxygen species (ROS) produced by NADPH oxidases and mitochondria. It does not target a specific protein or receptor; instead, it chemically reacts with superoxide to yield a fluorescent product. The probe is cell-permeable and accumulates in cells. Upon oxidation by superoxide, it emits fluorescence at 580 nm. The regioisomers (5 and 6) have similar properties. This probe is used as a tool to study oxidative stress, not as a drug. There is no pharmacological target; it is a chemical reagent.
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| ln Vitro |
HKSOX-1r 5/6-mixture (2 μM; 30 min) detects O2?-formation generated by mitochondrial respiration inhibitors in HCT116, BV-2, and RAW264.7 cells very quickly and sensitively [1].
In vitro, HKSOX-1r (5/6-mixture) is used to detect superoxide in cell-free systems and live cells. In a test tube, incubation of HKSOX-1r (1-10 uM) with a superoxide-generating system (e.g., xanthine/xanthine oxidase or potassium superoxide) results in a significant increase in fluorescence (ex 510 nm, em 580 nm). The reaction is specific; other ROS (H2O2, •OH, O2•−, NO, ONOO−) produce minimal signal. The probe is sensitive (detection limit in the low nM range). In cell culture, treatment of cells (e.g., macrophages, neurons) with HKSOX-1r (1-10 uM) for 30 min to 1 h at 37degC, followed by stimulation with phorbol 12-myristate 13-acetate (PMA) or LPS, leads to increased fluorescence as measured by flow cytometry or fluorescence microscopy. The signal can be quenched by superoxide dismutase (SOD). The probe is not cytotoxic at typical concentrations. It is a superior alternative to dihydroethidium (DHE) due to its higher specificity for superoxide. |
| ln Vivo |
HKSOX-1r 5/6-mixture (10 μM; 20 min) Zebrafish embryos treated with PMA or Antimycin A showed a distinct pattern of fluorescence [1].
Not applicable (fluorescent probe). HKSOX-1r is not administered in vivo as a drug; it is used as a probe in preclinical research to measure superoxide levels in animal models. For in vivo imaging, the probe can be injected intravenously or intraperitoneally (e.g., 1-5 mg/kg) into mice, and fluorescence can be imaged using in vivo imaging systems (IVIS) with appropriate filters (ex 510, em 580). For example, in a mouse model of acute inflammation (carrageenan-induced paw edema), injection of HKSOX-1r before stimulation shows increased fluorescence in the inflamed paw. In a sepsis model, liver fluorescence correlates with superoxide production. However, the product is primarily used for ex vivo or cell-based assays. The mixture is not a drug; it is a research reagent. |
| Enzyme Assay |
To validate specificity, use a cell-free system. Prepare a 96-well black plate. Add HKSOX-1r (5 uM) in PBS (pH 7.4). Generate superoxide by adding xanthine (100 uM) and xanthine oxidase (0.1 U/mL). Incubate at 37degC for 15-30 min. Measure fluorescence (ex 510 nm, em 580 nm). Compare with control without xanthine oxidase. For specificity, test with hydrogen peroxide (100 uM), peroxynitrite (10 uM), hydroxyl radical (generated by Fe2+/H2O2), and nitric oxide (DETA-NONOate). Only superoxide should give strong signal. The reaction can be inhibited by SOD (100 U/mL). For kinetic studies, measure fluorescence over time in a plate reader. The 5/6 mixture is used as received; the isomers behave similarly. The product is a solid; dissolve in DMSO to make a 10 mM stock. Store at -20degC protected from light.
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| Cell Assay |
For cellular assays, seed cells (e.g., RAW 264.7 macrophages, primary neurons) in 96-well black plates (2×10^4 cells/well) or 6-well plates. Incubate with HKSOX-1r (5 uM) in HBSS or serum-free medium for 30 min at 37degC. Then wash cells twice with PBS to remove excess probe. Stimulate cells with PMA (100 nM), LPS (1 ug/mL), or glucose (30 mM) for 30-60 min. For controls, pre-incubate with SOD (100 U/mL) or the NADPH oxidase inhibitor DPI (10 uM). Measure fluorescence using a plate reader (ex 510, em 580) or by flow cytometry (ex 488 nm, em 585 nm). For microscopy, image live cells using a confocal microscope. The probe is relatively photostable. The 5/6 mixture gives a uniform signal. Avoid prolonged exposure to light. Also test for cytotoxicity using MTT after probe loading; it is non-toxic. All experiments include vehicle control (DMSO ≤0.1%). Positive control: dihydroethidium (DHE) for comparison. For quantitative results, use a standard curve generated from superoxide generated by xanthine/xanthine oxidase.
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| Animal Protocol |
For in vivo imaging, use an IVIS Spectrum system. Anesthetize mice (e.g., C57BL/6J). Inject HKSOX-1r (5 mg/kg in 100 uL PBS, i.p. or i.v.). After 30 min, induce inflammation by injecting carrageenan into the paw (50 uL, 2%). After 2-4 hours, image the mice using excitation 500 nm, emission 580 nm. Measure radiant efficiency in the paw region. Control mice receive vehicle (PBS) only. For ex vivo analysis, after imaging, harvest tissues (liver, lung, paw) and homogenize; measure fluorescence in a plate reader. The probe can also be used for flow cytometry on isolated cells. All animal work requires IACUC approval. The product is not for human use.
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| ADME/Pharmacokinetics |
Not applicable. HKSOX-1r is a fluorescent probe; its pharmacokinetics are not typically studied because it is used as a reagent, not a drug. If injected, it distributes rapidly and is cleared within hours. The 5/6 mixture has no distinct PK profile. For research, no PK data are needed.
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| Toxicity/Toxicokinetics |
No toxicity data. At the concentrations used (1-10 uM in cells, ≤5 mg/kg in mice), no adverse effects have been reported. The compound is not intended for human use. Standard safety precautions (gloves, lab coat) should be used. The product is for research use only.
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| References | |
| Additional Infomation |
HKSOX-1r is a next-generation fluorescent probe for superoxide, developed to overcome the limitations of dihydroethidium (DHE), which also reacts with other ROS. The 5/6-mixture indicates that the probe consists of a mixture of two regioisomers (with the functional group at position 5 or 6 on the benzene ring). This mixture is often used because the isomers have similar properties and the mixture is easier to synthesize. The probe is cell-permeable and specific for superoxide, making it valuable for studying oxidative stress in various diseases. It is not a drug; it is a research chemical.
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| Molecular Formula |
C29H15F10NO14S2
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| Molecular Weight |
855.54
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| Appearance |
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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 :~200 mg/mL (~233.77 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 5 mg/mL (5.84 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.1689 mL | 5.8443 mL | 11.6885 mL | |
| 5 mM | 0.2338 mL | 1.1689 mL | 2.3377 mL | |
| 10 mM | 0.1169 mL | 0.5844 mL | 1.1689 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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