| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
Peroxynitrite (ONOO-) is the primary molecular target. The probe is rationally designed at the University of Hong Kong for the selective detection of ONOO- over other reactive oxygen species (ROS).
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|---|---|
| ln Vitro |
1. The working solution for HKYellow-AM (6/12-mixture) is prepared as follows: dissolve 1 mg of HKYellow-AM (6/12-mixture) in 70 μL DMSO to create a 10 mM stock solution. Note: To prevent frequent freezing and thawing, it is advised that the stock solution be kept between -20°C and 80°C in the dark. 1.2 HKYellow-AM (6/12-mixture) working solution To create a 1–10 μM working solution, dilute the stock solution with either PBS or serum-free cell culture medium. Note: Please modify the working solution concentration of HKYellow-AM (6/12-mixed solution) based on the actual circumstances. 2. 2.1 Suspension cells (six-well plate) 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.
HKYellow-AM exhibits high selectivity and sensitivity for ONOO- in cell-free assays. Upon reaction with ONOO-, the probe generates a strong fluorescence turn-on signal. The mixture consists of two components (6/12-positional isomers) that exhibit similar spectroscopic properties. |
| 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 ONOO- in live cells and animal models.
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| Enzyme Assay |
A standard protocol involves incubating HKYellow-AM (e.g., 10 uM) with varying concentrations of ONOO- in a buffer solution (e.g., PBS, pH 7.4) for 5-10 minutes at room temperature. The fluorescence intensity is then measured using a spectrophotometer with appropriate excitation and emission wavelengths.
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| Cell Assay |
A general protocol involves seeding cells in a confocal dish and incubating with HKYellow-AM (e.g., 5 uM) for 20-30 minutes at 37degC. The AM ester group facilitates cell permeability. After incubation, cells are washed and fluorescence imaging is performed using a confocal microscope.
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| Animal Protocol |
A typical protocol for in vivo imaging involves intravenous or intraperitoneal injection of an optimized concentration of HKYellow-AM 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 ONOO--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 HKYellow-AM, the product is reported to have no cytotoxicity at working concentrations. 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 | |
| Additional Infomation |
HKYellow-AM is derived from patent EP2809666B1 (compound 14). The AM ester form enhances cell permeability, allowing it to cross cell membranes. Once inside cells, esterases cleave the AM group, trapping the active HKYellow probe inside for imaging purposes.
|
| Molecular Formula |
C40H39CLN2O8
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|---|---|
| Molecular Weight |
711.199270486832
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| Exact Mass |
710.239
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| CAS # |
1448821-89-3
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| PubChem CID |
162640791
|
| Appearance |
Purple to purplish red solid powder
|
| LogP |
7.7
|
| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
51
|
| Complexity |
1290
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| Defined Atom Stereocenter Count |
0
|
| SMILES |
O=C1C2=CC=CC=C2C2(C3=CC(Cl)=C(N(C4C=CC(O)=CC=4)C)C=C3OC3=CC4N(C(C)(C)CC(C)C=4C=C23)CCCC(=O)OCOC(=O)C)O1
|
| InChi Key |
AIRAPNPEUVLNDG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C40H39ClN2O8/c1-23-21-39(3,4)43(16-8-11-37(46)49-22-48-24(2)44)33-19-35-30(17-28(23)33)40(29-10-7-6-9-27(29)38(47)51-40)31-18-32(41)34(20-36(31)50-35)42(5)25-12-14-26(45)15-13-25/h6-7,9-10,12-15,17-20,23,45H,8,11,16,21-22H2,1-5H3
|
| Chemical Name |
acetyloxymethyl 4-[8'-chloro-9'-(4-hydroxy-N-methylanilino)-2',2',4'-trimethyl-3-oxospiro[2-benzofuran-1,6'-3,4-dihydrochromeno[3,2-g]quinoline]-1'-yl]butanoate
|
| 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: 130 mg/mL (91.39 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 3.25 mg/mL (2.28 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 32.5 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: 3.25 mg/mL (2.28 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 32.5 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 | 1.4061 mL | 7.0304 mL | 14.0607 mL | |
| 5 mM | 0.2812 mL | 1.4061 mL | 2.8121 mL | |
| 10 mM | 0.1406 mL | 0.7030 mL | 1.4061 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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