Lucigenin

• CAT Number: R019285
• CAS Number: 2315-97-1
• Molecular Formula: C28H22N4O6
• Molecular Weight: 510.50
• Purity: ≥95%
• Synonyms: 10-methyl-9-(10-methylacridin-10-ium-9-yl)acridin-10-ium;dinitrate
• InChI: 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
• InChIKey: KNJDBYZZKAZQNG-UHFFFAOYSA-N
• SMILES: C[N+]1=C2C=CC=CC2=C(C3=CC=CC=C31)C4=C5C=CC=CC5=[N+](C6=CC=CC=C64)C.[N+](=O)([O-])[O-].[N+](=O)([O-])[O-]

Lucigenin is a chemiluminescence probe. Lucigenin can be used to detect the production of endogenous superoxide anion radical (O2-). Lucigenin is extremely sensitive to chloride ions, while it combined with chloride ions, the fluorescence will be quenched. Lucigenin also can be used as a chloride indicator. Ex/Em=455/505 nm[1].
Preparation of Lucigenin working solution
1.1 Preparation of the stock solution
Dissolve 1 mg of Lucigenin in 0.1919 mL of DMSO to obtain 10 mM of Lucigenin.
Note: It is recommended to store the stock solution at -20 °C -80 °C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Lucigenin working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 5-10 μM of Lucigenin working solution.
Note: Please adjust the concentration of Lucigenin working solution according to the actual situation.
Cell staining
2.1 Cell preparation.For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of Lucigenin working solution, and then incubate at room temperature for 15 minutes.
2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope.

Reference

[1]. Li Y, et al. Validation of lucigenin (bis-N-methylacridinium) as a chemilumigenic probe for detecting superoxide anion radical production by enzymatic and cellular systems. J Biol Chem. 1998 Jan 23;273(4):2015-23.
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