| 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 highly selective detection of H2O2 over 30-fold other tested reactive oxygen/nitrogen species (ROS/RNS) in chemical and biological systems.
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|---|---|
| ln Vitro |
1. Working solution preparation for HKPerox-2 1.1 Stock solution preparation: Dissolve 1 milligram of HKPerox-2 in 172 uL DMF 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 HKPerox-2 working solution: Use serum-free cell culture medium to dilute the stock solution or PBS to create a working solution that ranges from 1 to 10 μM. It is important to modify the concentration of HKPerox-2 working solution based on the specific circumstances. 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.
HKPerox-2 exhibits excellent selectivity and sensitivity for H2O2 in cell-free assays. Upon reaction with H2O2 via a tandem Payne/Dakin reaction, the probe undergoes a specific chemical transformation that generates a strong fluorescence turn-on signal. It can detect H2O2 at low concentrations. |
| ln Vivo |
There is no specific in vivo activity data available for this compound. As a highly selective green fluorescent probe, HKPerox-2 can be used for real-time imaging of endogenous and exogenous H2O2 in live cells and potentially in small animal models.
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| Enzyme Assay |
A standard protocol involves incubating HKPerox-2 (e.g., 5 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 (e.g., ·OH, O2-, NO, ONOO-).
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| Cell Assay |
A general protocol for cellular imaging involves seeding cells in a confocal dish and incubating with HKPerox-2 (e.g., 1-10 uM) for 30 minutes at 37degC. Cells are then washed to remove excess probe and treated with an H2O2 donor or an oxidative stress inducer (e.g., PMA or rotenone). Fluorescence imaging is performed with a confocal microscope using 520 nm excitation and 543 nm emission collection.
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| Animal Protocol |
A typical protocol for in vivo imaging involves intravenous or intraperitoneal injection of an optimized concentration of HKPerox-2 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, as these compounds are typically used for acute imaging. For in vivo use, HKPerox-2 can be formulated in PBS with 1-5% DMSO or in saline.
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| Toxicity/Toxicokinetics |
General toxicity for imaging probes is considered low. No specific toxicity data is available for HKPerox-2. For similar compounds like HKPerox-1, incubating with up to 20 uM for 24 hours in RAW264.7 macrophages shows no cytotoxicity.
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| References | |
| Additional Infomation |
HKPerox-2 is a research tool for studying the role of H2O2 in oxidative stress, signaling, and various diseases such as inflammation, neurodegeneration, and cancer. Its high selectivity over other ROS/RNS makes it a valuable reagent for dissecting H2O2-specific pathways.
|
| Molecular Formula |
C32H23NO10
|
|---|---|
| Molecular Weight |
581.53
|
| Exact Mass |
1178.296
|
| CAS # |
2245230-79-7
|
| PubChem CID |
168325903
|
| Appearance |
Yellow to orange solid powder
|
| Hydrogen Bond Donor Count |
4
|
| Rotatable Bond Count |
14
|
| Heavy Atom Count |
87
|
| Complexity |
2170
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C.CN(C1=CC2=C(C=C1)C3(C4=C(C=C(C=C4)C(=O)O)C(=O)O3)C5=C(O2)C=C(C=C5)OC)C(=O)OCC6=CC(=C(C=C6)C=O)O.CN(C1=CC2=C(C=C1)C3(C4=C(O2)C=C(C=C4)OC)C5=C(C=CC(=C5)C(=O)O)C(=O)O3)C(=O)OCC6=CC(=C(C=C6)C=O)O
|
| InChi Key |
PEPBXYWAYSPIJB-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/2C32H23NO10.CH4/c1-33(31(39)41-16-17-3-4-19(15-34)26(35)11-17)20-6-9-24-27(13-20)42-28-14-21(40-2)7-10-25(28)32(24)23-8-5-18(29(36)37)12-22(23)30(38)43-32;1-33(31(39)41-16-17-3-4-19(15-34)26(35)11-17)20-6-9-23-27(13-20)42-28-14-21(40-2)7-10-24(28)32(23)25-12-18(29(36)37)5-8-22(25)30(38)43-32;/h2*3-15,35H,16H2,1-2H3,(H,36,37);1H4
|
| Chemical Name |
3'-[(4-formyl-3-hydroxyphenyl)methoxycarbonyl-methylamino]-6'-methoxy-1-oxospiro[2-benzofuran-3,9'-xanthene]-5-carboxylic acid;3'-[(4-formyl-3-hydroxyphenyl)methoxycarbonyl-methylamino]-6'-methoxy-3-oxospiro[2-benzofuran-1,9'-xanthene]-5-carboxylic acid;methane
|
| 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 (85.98 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 | 1.7196 mL | 8.5980 mL | 17.1960 mL | |
| 5 mM | 0.3439 mL | 1.7196 mL | 3.4392 mL | |
| 10 mM | 0.1720 mL | 0.8598 mL | 1.7196 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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