| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
Hydroxyl radicals (·OH) are the primary molecular target. The probe is designed for the selective detection of ·OH over other reactive oxygen species (ROS).
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|---|---|
| ln Vitro |
1. Preparation of HKOH-1 working solution 1.1 Get the stock solution ready. To get a 10 mM stock solution, dilute 1 milligram of HKOH-1 with 135 μL of anhydrous DMSO. It is advised to aliquot and store HKOH-1 storage solution at -20°C or -80°C in the dark. 1.2 Creating a functional solution Use preheated serum-free cell culture medium or PBS dilution stock solution to prepare a 1-10 μM HKOH-1 working solution. Note: Please prepare the HKOH-1 working fluid for usage by adjusting the concentration to suit the current circumstances. 2. Cell staining (suspension cells) 2.1 Centrifuge the cells, add PBS, and wash twice for five minutes each time. The density of cells is 1×106/mL. 2.2 Add 1 mL of the HKOH-1 working solution, then let it sit at room temperature for five to thirty minutes. 2.3 Centrifuge for 3–4 minutes at 400 g, then remove the supernatant. 2.4 After adding PBS, wash the cells twice for five minutes each. 2.5 Re-suspend the cells in 1 milliliter of PBS or serum-free media, and use a flow cytometer or fluorescence microscope to observe. 3. Staining adherent cells in cells 3.1 Use sterile coverslips to cultivate adherent cells. 3.2 Aspirate extra culture medium after removing the coverslip from the medium. 3.3 Cover the cells fully with 100 μL of the dye working solution, shake gently, and allow incubate for five to thirty minutes. 3.4 Use a flow cytometer or fluorescence microscope to observe after aspirating the dye working solution and washing it twice in culture media for a total of five minutes each time.
HKOH-1 is a highly sensitive and selective fluorescent probe for detecting endogenous hydroxyl radicals. It shows absorption peaks at 500 nm and exhibits a strong fluorescence turn-on signal in the presence of ·OH. Its excitation/emission maxima are 500/520 nm. |
| ln Vivo |
There is no specific in vivo activity data available for this compound. However, as a fluorescent probe for reactive species, similar compounds are typically used for real-time imaging of ·OH in live cells and animal models.
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| Enzyme Assay |
A standard protocol involves incubating HKOH-1 (e.g., 5 uM) with varying concentrations of hydroxyl radical-generating systems in a buffer solution (e.g., PBS, pH 7.4) at room temperature. The fluorescence intensity (Ex/Em: 500/520 nm) is then measured with a spectrophotometer.
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| Cell Assay |
A general protocol for cellular imaging involves seeding cells in a confocal dish and incubating with HKOH-1 (e.g., 1-10 uM) for 20-30 minutes at 37degC. After washing, the cells are treated with an oxidative stress inducer to generate endogenous ·OH, and fluorescence is imaged with a confocal microscope (Ex/Em: 500/520 nm).
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| Animal Protocol |
A typical protocol for in vivo imaging involves intravenous or intraperitoneal injection of an optimized concentration of HKOH-1 into a live animal model. After a suitable circulation time (e.g., 30 minutes to 2 hours), the animal is placed under an in vivo imaging system to monitor ·OH-induced fluorescence.
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| ADME/Pharmacokinetics |
General PK properties for small molecule probes are not applicable, as these compounds are typically used for acute imaging. However, formulation can be in PBS or saline with co-solvents like DMSO for in vivo use.
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| Toxicity/Toxicokinetics |
General toxicity for imaging probes is considered low. For HKOH-1, no specific toxicity studies are reported. In similar experiments, incubation with up to 10 uM for 24 hours in cell models shows good biocompatibility and no significant cytotoxicity.
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| References |
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| Additional Infomation |
HKOH-1 is a research tool for studying oxidative stress and related diseases. The probe's turn-on response offers a low background, making it ideal for high-contrast imaging of hydroxyl radicals. This product is for research use only and not for diagnostic or therapeutic applications.
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| Molecular Formula |
C26H12CL2I2O6
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|---|---|
| Molecular Weight |
745.084830284119
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| Exact Mass |
743.81
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| CAS # |
2031170-96-2
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| PubChem CID |
122647002
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| Appearance |
White to off-white solid powder
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| LogP |
7.5
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
36
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| Complexity |
841
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C12(C3=C(C=C(OC4=CC(I)=C(O)C(I)=C4)C(Cl)=C3)OC3=C1C=C(Cl)C(O)=C3)C1=C(C=CC=C1)C(=O)O2
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| InChi Key |
BZWBSIJSVLAKAF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H12Cl2I2O6/c27-16-7-14-21(9-20(16)31)35-22-10-23(34-11-5-18(29)24(32)19(30)6-11)17(28)8-15(22)26(14)13-4-2-1-3-12(13)25(33)36-26/h1-10,31-32H
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| Chemical Name |
2',7'-dichloro-3'-hydroxy-6'-(4-hydroxy-3,5-diiodophenoxy)spiro[2-benzofuran-3,9'-xanthene]-1-one
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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: 180 mg/mL (241.58 mM)
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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 | 1.3421 mL | 6.7107 mL | 13.4214 mL | |
| 5 mM | 0.2684 mL | 1.3421 mL | 2.6843 mL | |
| 10 mM | 0.1342 mL | 0.6711 mL | 1.3421 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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