| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
|
||
| 250mg |
|
||
| 500mg |
|
||
| Other Sizes |
| Targets |
Lucigenin targets superoxide anion radical (O₂⁻) as a chemilumigenic probe[1]
|
|---|---|
| ln Vitro |
1.1 Making the stock solution: To get 10 mM lucigenin, dissolve 1 milligram of lucigenin in 0.1919 mL of DMSO. Note: To prevent frequent freezing and thawing, it is advised to store the stock solution at -20°C or -80°C away from light. 1.2 Lucigenin working solution: To create a 5–10 μM Lucigenin working solution, dilute the original solution with serum-free cell solution or PBS. Note: Please modify the Lucigenin working solution concentration based on the actual circumstances. 2.1 Preparing suspended cells for staining: Centrifuge at 1000 g for 3–5 minutes at 4°C; discard supernatant. Use PBS to wash twice, for five minutes each time. Adherent cells: To create a single cell suspension, add islet dissociated cells and discard the cell culture media. Discard the supernatant after centrifuging at 1000g for three to five minutes at 4°C. Use PBS to wash twice, for five minutes each time. 2.2 After adding 1 milliliter of the Lucigenin working solution, season for 15 minutes. 2.3 Discard the supernatant after centrifuging at 400 g for three to four minutes at 4°C. 2.4 Use PBS twice, giving each wash five minutes. 2.5 Re-suspend the cells in PBS or serum-free cells and use a fluorescent microscope to observe.
- Enzymatic system superoxide anion detection: Lucigenin produced dose-dependent chemiluminescence in the xanthine oxidase-xanthine system, with a linear response to O₂⁻ concentrations ranging from 0.1 to 10 nmol/L. The chemiluminescence signal was significantly inhibited (≥ 90%) by superoxide dismutase (SOD), confirming specificity for O₂⁻ [1] - Cellular system superoxide anion detection: Lucigenin detected O₂⁻ production in phorbol 12-myristate 13-acetate (PMA)-stimulated RAW 264.7 macrophages. The chemiluminescence signal peaked at 15-20 minutes after stimulation and was reduced by 85% in the presence of SOD. Unstimulated macrophages showed minimal chemiluminescence, indicating low background noise [1] - Selectivity verification: Lucigenin did not produce significant chemiluminescence with other reactive oxygen species (ROS) including hydrogen peroxide (H₂O₂), hydroxyl radical (·OH), and singlet oxygen (¹O₂), confirming high selectivity for O₂⁻ [1] |
| Enzyme Assay |
- Xanthine oxidase-xanthine system chemiluminescence assay: Reaction mixtures were prepared by mixing xanthine (final concentration 0.1 mM), xanthine oxidase (0.01 U/mL), and Lucigenin (final concentration 5 μM) in phosphate-buffered saline (pH 7.4). For selectivity testing, SOD (100 U/mL) was added to parallel mixtures. Chemiluminescence intensity was measured continuously for 30 minutes using a chemiluminometer. The linear range of O₂⁻ detection was determined by varying xanthine oxidase concentration to adjust O₂⁻ production [1]
- Reactive oxygen species (ROS) selectivity assay: Separate reaction mixtures containing Lucigenin (5 μM) and individual ROS generators (H₂O₂: 1 mM; ·OH: generated by Fenton reaction; ¹O₂: generated by hypochlorite-hydrogen peroxide system) were prepared. Chemiluminescence was measured for 30 minutes to evaluate cross-reactivity with non-target ROS [1] |
| Cell Assay |
- Macrophage superoxide anion detection assay: RAW 264.7 macrophages were seeded into 96-well plates at a density of 5×10⁴ cells/well and incubated overnight. The medium was replaced with serum-free medium containing Lucigenin (5 μM), and the cells were pre-incubated for 30 minutes. PMA (final concentration 100 nM) was added to stimulate O₂⁻ production, while unstimulated cells served as the negative control. For specificity verification, SOD (100 U/mL) was added to parallel wells. Chemiluminescence intensity was measured every 5 minutes for 60 minutes using a microplate chemiluminometer [1]
|
| References | |
| Additional Infomation |
See also: Rushikin (note moved to).
- Chemical properties: Luxigin (bis-N-methylacridine) is a cationic organic compound belonging to the acridine salt family. It is characterized by its ability to react with superoxide anion free radicals in a chemiluminescent reaction [1] - Mechanism of action: Luxigin reacts with O₂⁻ through single-electron reduction to generate free radical cations, which then decompose and emit light (maximum emission wavelength is about 480 nm), thus enabling quantitative detection of O₂⁻ [1] - Probe advantages: Luxigin has high sensitivity to O₂⁻ (detection limit is about 0.05 nmol/L), low background chemiluminescence, and is compatible with both enzyme and cell systems, making it a reliable tool for O₂⁻ research [1] - Key verification: Specific luminol detection of O₂⁻ depends on SOD inhibition—only O₂⁻ The mediated chemiluminescence is suppressed by SOD, which distinguishes it from other ROS probes [1] |
| Molecular Formula |
C28H22N4O6
|
|---|---|
| Molecular Weight |
510.5
|
| Exact Mass |
510.153
|
| CAS # |
2315-97-1
|
| PubChem CID |
65099
|
| Appearance |
Yellow to khaki solid powder
|
| Melting Point |
250ºC
|
| LogP |
6.183
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
38
|
| Complexity |
522
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
KNJDBYZZKAZQNG-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C28H22N2.2NO3/c1-29-23-15-7-3-11-19(23)27(20-12-4-8-16-24(20)29)28-21-13-5-9-17-25(21)30(2)26-18-10-6-14-22(26)28;2*2-1(3)4/h3-18H,1-2H3;;/q+2;2*-1
|
| Chemical Name |
10-methyl-9-(10-methylacridin-10-ium-9-yl)acridin-10-ium;dinitrate
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~62.5 mg/mL (~122.43 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.07 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 20.8 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: ≥ 2.08 mg/mL (4.07 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (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 20.8 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 | 1.9589 mL | 9.7943 mL | 19.5886 mL | |
| 5 mM | 0.3918 mL | 1.9589 mL | 3.9177 mL | |
| 10 mM | 0.1959 mL | 0.9794 mL | 1.9589 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
