| 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, a highly reactive oxygen species, over other ROS.
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|---|---|
| ln Vitro |
1. Working solution preparation for HKOH-1r 1.1: Prepare the stock solution by dissolving 1 milligram of HKOH-1r in 107 μL DMSO to create a 10 mM stock solution. Note: It is advised to keep the stock solution out of direct sunlight and between -20°C and 80°C to prevent repeated freezing and thawing. 1.2 Making the HKOH-1r working solution: Use serum-free cell culture medium to dilute the stock solution or PBS to create a working solution that is 1–10 μM. Please note: Based on the current circumstances, modify the HKOH-1r working fluid concentration. 2. Suspension cells in a 6-well plate (2.1) are stained A. After centrifuging at 1000g for three to five minutes at 4°C, discard the supernatant. Wash for five minutes each time, twice, using PBS. 1×106/mL b is the cell density. After adding 1 mL of the working solution, let it sit at room temperature for five to thirty minutes. an. After centrifuging at 400 g for three to four minutes at 4 °C, discard the supernatant. D. Wash for five minutes each time, twice, using PBS. E. Using a fluorescence microscope or flow cytometry, resuspend the cells in PBS or serum-free cell culture media. Cells that adhere 2.2 a. Adherent cells are cultured on sterile coverslips. B. Lift the coverslip out of the culture media and use the aspirator to remove any extra. an. To fully cover the cells, add 100 μL of working solution, shake gently, and then incubate for five to thirty minutes at room temperature. D. Wash for five minutes twice using a medium setting. either flow cytometry monitoring or fluorescence microscopy.
HKOH-1r exhibits high sensitivity and selectivity for hydroxyl radicals in cell-free assays. Upon reaction with ·OH, the probe undergoes a chemical transformation that results in a strong fluorescence turn-on or ratiometric signal, enabling accurate detection. |
| ln Vivo |
There is no specific in vivo activity data available for this compound. As a fluorescent probe for reactive species, it is designed for use in live cells, and similar compounds are typically used for imaging ·OH in animal models as well.
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| Enzyme Assay |
A standard protocol for testing ·OH detection involves incubating HKOH-1r (e.g., 5 uM) with varying concentrations of a hydroxyl radical-generating system (e.g., Fenton reaction: Fe2+ + H2O2) in a buffer solution (e.g., PBS, pH 7.4) at room temperature. The fluorescence intensity ratio (e.g., 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 (e.g., macrophages or neuronal cells) in a confocal dish and incubating with HKOH-1r (e.g., 1-10 uM) for 20-30 minutes at 37degC. Cells are then washed to remove excess probe. For ratiometric imaging, cells are treated with an oxidative stress inducer to generate ·OH, and images at two emission wavelengths are captured and processed to obtain ratio images.
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| Animal Protocol |
A typical protocol for in vivo imaging involves intravenous or intraperitoneal injection of an optimized concentration of HKOH-1r 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-1r, no specific toxicity studies are reported. In similar experiments with related probes (e.g., HKPerox-1 at 1-20 uM for 24 h in RAW264.7 macrophages), no cytotoxicity was observed.
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| References |
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| Additional Infomation |
HKOH-1r is a research tool for studying oxidative stress and hydroxyl radical-mediated damage in a variety of biological contexts, including neuroinflammation, ischemia-reperfusion injury, and aging. The ratiometric design allows for more accurate quantification of ·OH levels compared to turn-on-only probes.
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| Molecular Formula |
C34H25CL2I2NO11
|
|---|---|
| Molecular Weight |
948.2774
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| Exact Mass |
946.889
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| CAS # |
2138472-08-7
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| PubChem CID |
156588681
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| Appearance |
White to yellow solid powder
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
50
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| Complexity |
1060
|
| Defined Atom Stereocenter Count |
0
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| SMILES |
IC1C(=C(C([H])=C(C=1[H])OC1=C(C([H])=C2C(=C1[H])OC1=C([H])C(=C(C([H])=C1C12C2=C([H])C([H])=C([H])C([H])=C2C(=O)O1)Cl)O[H])Cl)I)O[H].O(C([H])([H])[H])C(C([H])([H])N(C(C([H])([H])[H])=O)C([H])([H])C(=O)OC([H])([H])[H])=O
|
| InChi Key |
VHPOZCLVPZPUPA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H12Cl2I2O6.C8H13NO5/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;1-6(10)9(4-7(11)13-2)5-8(12)14-3/h1-10,31-32H;4-5H2,1-3H3
|
| Chemical Name |
2',7'-dichloro-3'-hydroxy-6'-(4-hydroxy-3,5-diiodophenoxy)spiro[2-benzofuran-3,9'-xanthene]-1-one;methyl 2-[acetyl-(2-methoxy-2-oxoethyl)amino]acetate
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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: 95 mg/mL (101.25 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 4.75 mg/mL (5.06 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 47.5 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: 4.75 mg/mL (5.06 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 47.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.0545 mL | 5.2727 mL | 10.5454 mL | |
| 5 mM | 0.2109 mL | 1.0545 mL | 2.1091 mL | |
| 10 mM | 0.1055 mL | 0.5273 mL | 1.0545 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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