| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
Hydrogen peroxide (H2O2) is the primary molecular target. The probe is designed for the selective detection of H2O2 with excellent selectivity and sensitivity, enabling molecular imaging of endogenous H2O2 in living cells.
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|---|---|
| ln Vitro |
1. 1.1 Creating the working solution for HKPerox-1 Dilute 1 milligram of HKPerox-1 with 140 μL of anhydrous DMF to create a 10 mM stock solution. It is advised that HKPerox-1 storage solution be aliquoted and kept in the dark at -20°C or -80°C. 1.2 Making the working solution: To make a 1-10 μM HKPerox-1 working solution, use warmed serum-free cell culture medium or PBS dilution stock solution. Note: Please be ready for instant use by adjusting the HKPerox-1 working fluid concentration based on the current circumstances. 2. Staining cells (suspension cells) 2.1 Centrifuge the cells, add PBS, then wash twice for five minutes each. One x 10^6 cells per milliliter. 2.2 After adding 1 mL of HKPerox-1 working solution, let it sit at room temperature for a duration of 5 to 30 minutes. 2.3 Discard the supernatant after centrifuging for 3–4 minutes at 400 g. 2.4 Add PBS and give the cells two 5-minute washing cycles. 2.5 Use a flow cytometer or fluorescence microscope to view the cells after resuspending them in 1 mL of serum-free media or PBS. 3. 3. Culture adherent cells on sterile coverslips for cell labeling (adherent cells). 3.2 Take off the coverslip from the culture media and use an aspirator to remove any extra. 3.3 Add 100 μL of the dye working solution, give the cells a gentle shake to cover them completely, and then let them sit for five to thirty minutes. 3.4 Aspirate the dye working solution, rinse with culture medium two or three times for five minutes each time, and use a flow cytometer or fluorescence microscope to monitor.
In cell-free assays, HKPerox-1 exhibits excellent selectivity and sensitivity for H2O2 over other reactive oxygen species (ROS). Upon reaction with H2O2, the probe undergoes a specific chemical transformation that generates a strong fluorescence turn-on signal. |
| ln Vivo |
There is no specific in vivo activity data available for this compound. As a highly selective green fluorescent probe, HKPerox-1 is used for real-time imaging of endogenous H2O2 in live cells. Similar probes are also used in small animal models for imaging oxidative stress.
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| Enzyme Assay |
A standard protocol involves incubating HKPerox-1 (e.g., 5-10 uM) with varying concentrations of H2O2 in a buffer solution (e.g., PBS, pH 7.4) for 5-30 minutes at 37degC. The fluorescence intensity (Ex/Em: 520/543 nm) is then measured with a fluorescence spectrophotometer. Selectivity is confirmed by testing against other ROS/RNS.
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| Cell Assay |
A general protocol for cellular imaging involves seeding cells (e.g., RAW264.7 macrophages) in a confocal dish and incubating with HKPerox-1 (e.g., 1-20 uM) for 30 minutes at 37degC. Cells are then washed to remove excess probe and treated with an H2O2 donor (e.g., 50-200 uM H2O2) or an oxidative stress inducer (e.g., phorbol myristate acetate, PMA). Fluorescence imaging is performed with a confocal microscope. A control group can be pretreated with an antioxidant (e.g., N-acetylcysteine) to suppress signal.
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| Animal Protocol |
A typical protocol for in vivo imaging involves intravenous or intraperitoneal injection of an optimized concentration of HKPerox-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 H2O2-induced fluorescence.
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| ADME/Pharmacokinetics |
General PK properties for small molecule probes are not applicable. For in vivo use, HKPerox-1 can be formulated in PBS with 1-5% DMSO or in saline. For cellular experiments, stock solutions are typically prepared in DMSO (e.g., 10 mM).
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| Toxicity/Toxicokinetics |
General toxicity for imaging probes is considered low. HKPerox-1 (1-20 uM; 24 hours) is reported to be nontoxic to RAW264.7 macrophages. This indicates good biocompatibility at working concentrations.
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| References | |
| Additional Infomation |
HKPerox-1 is a research tool for studying the role of H2O2 in oxidative stress, redox signaling, inflammation, and various diseases including cancer, neurodegeneration, and cardiovascular disorders. Its high sensitivity allows detection of endogenous H2O2 produced by cells under normal physiological conditions or in response to stimuli.
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| Molecular Formula |
C40H30N2O11
|
|---|---|
| Molecular Weight |
714.673011302948
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| Exact Mass |
714.184
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| CAS # |
2240163-05-5
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| PubChem CID |
162640794
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| Appearance |
Yellow to orange solid powder
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| LogP |
6
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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 |
10
|
| Heavy Atom Count |
53
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| Complexity |
1280
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O=C1C2=CC=CC=C2C2(C3=CC=C(N(C)C(=O)OCC4C=CC(C=O)=C(O)C=4)C=C3OC3=CC(N(C)C(=O)OCC4C=CC(C=O)=C(O)C=4)=CC=C23)O1
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| InChi Key |
WOSPPMBMPHNEFP-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C40H30N2O11/c1-41(38(48)50-21-23-7-9-25(19-43)33(45)15-23)27-11-13-31-35(17-27)52-36-18-28(42(2)39(49)51-22-24-8-10-26(20-44)34(46)16-24)12-14-32(36)40(31)30-6-4-3-5-29(30)37(47)53-40/h3-20,45-46H,21-22H2,1-2H3
|
| Chemical Name |
(4-formyl-3-hydroxyphenyl)methyl N-[6'-[(4-formyl-3-hydroxyphenyl)methoxycarbonyl-methylamino]-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl]-N-methylcarbamate
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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) |
DMF: 50 mg/mL (69.96 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (3.50 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 25.0 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: ≥ 2.5 mg/mL (3.50 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.= View More
Solubility in Formulation 3: 1.25 mg/mL (1.75 mM) in 10% DMF 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Solubility in Formulation 4: ≥ 1.25 mg/mL (1.75 mM) (saturation unknown) in 10% DMF 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3992 mL | 6.9962 mL | 13.9925 mL | |
| 5 mM | 0.2798 mL | 1.3992 mL | 2.7985 mL | |
| 10 mM | 0.1399 mL | 0.6996 mL | 1.3992 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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