| 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-1 is a chemoselective fluorescent probe that specifically reacts with superoxide via a chemical mechanism (likely a reaction with the dihydroethidine core). The reaction yields a fluorescent product (ethidium-like). The 5/6 mixture indicates that the probe is a mixture of isomers where the substituent is at the 5 or 6 position on the benzene ring; both have similar properties. This probe has high selectivity for superoxide over other ROS (H2O2, •OH, ONOO−, NO). It is used in biochemistry and cell biology to monitor superoxide production from sources such as NADPH oxidases and mitochondria. There is no receptor target.
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| ln Vitro |
In potassium phosphate buffer at 25 °C, HKSOX-1 (10 μM) combines with O2?-to produce a strong time-dependent fluorescence increase that lasts for 60 minutes without changing in intensity. The reaction is finished in 10 minutes. HKSOX-1 (10 μM) has remarkable stability against pH variations throughout the 2.2–8.8 range [1].
In vitro, HKSOX-1 (5/6-mixture) (1-10 uM) reacts with superoxide generated by xanthine/xanthine oxidase, producing a fluorescence increase. The reaction is specific: no significant fluorescence is observed with H2O2, peroxynitrite, nitric oxide, or hydroxyl radical. The probe is sensitive (detection limit in the low nM range). In cell culture, loading cells with HKSOX-1 (5 uM) for 30 min, then stimulating with PMA or LPS, results in increased fluorescence measured by flow cytometry or plate reader. The signal is inhibited by superoxide dismutase (SOD) or NADPH oxidase inhibitors (e.g., DPI). The 5/6 mixture is used as supplied; the isomers perform similarly. The probe is not cytotoxic at 5 uM for 4 h. It is an improvement over dihydroethidium (DHE) due to better specificity. |
| ln Vivo |
Not applicable. HKSOX-1 is not a drug; it is a fluorescent probe. It can be used in ex vivo tissues or in vivo imaging. For in vivo studies, inject the probe intravenously (1-5 mg/kg) into mice; fluorescence can be imaged in organs with an IVIS system. For example, in a model of sepsis, liver fluorescence correlates with superoxide production. However, the mixture is primarily a research reagent.
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| Enzyme Assay |
Same as for HKSOX-1r and HKSOX-1m. In a cell-free system, dissolve HKSOX-1 in PBS (5 uM). Add xanthine (100 uM) and xanthine oxidase (0.1 U/mL). Incubate at 37degC for 20 min. Measure fluorescence (ex 510, em 580). Specificity confirmed by lack of signal with other ROS and inhibition by SOD. The 5/6 mixture is used directly. Dissolve in DMSO to make a 10 mM stock, then dilute in PBS. The probe is light-sensitive; handle in low light.
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| Cell Assay |
Same as for HKSOX-1r. Seed RAW 264.7 cells in 96-well plates, load with HKSOX-1 (5 uM) in HBSS for 30 min, wash, stimulate with PMA (100 nM) or LPS (1 ug/mL) for 30-60 min. Read fluorescence. For flow cytometry, load and stimulate in suspension. For microscopy, use live-cell imaging. The 5/6 mixture works well in these assays. Include controls with DMSO and SOD. The probe is not cytotoxic.
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| Animal Protocol |
Same as for HKSOX-1r. For in vivo, inject HKSOX-1 (5 mg/kg, i.v.) into mice, then induce inflammation. Image using IVIS. All animal work requires IACUC approval.
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| Toxicity/Toxicokinetics |
No toxicity. Low concentrations used. Standard safety.
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| References | |
| Additional Infomation |
HKSOX-1 is the original superoxide probe of the HKSOX family. It is highly selective for superoxide over other reactive species. The 5/6 mixture simplifies synthesis. It is widely used in oxidative stress research. Not for human use.
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| Molecular Formula |
C23H6F10O11S2
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|---|---|
| Molecular Weight |
712.40
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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: 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) |
DMSO :~110 mg/mL (~154.41 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.75 mg/mL (3.86 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 27.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.75 mg/mL (3.86 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 27.5 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 | 1.4037 mL | 7.0185 mL | 14.0371 mL | |
| 5 mM | 0.2807 mL | 1.4037 mL | 2.8074 mL | |
| 10 mM | 0.1404 mL | 0.7019 mL | 1.4037 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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