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Liperfluo

Item # Unit Size
L248-10
1 set (50 ug x 5)

For Research Use Only Products

Application: lipid peroxidase detection

Lipid peroxide selective measurement
Less photo-damage to cells
Applicable for microscopy and FCM analysis
 MSDS
Chemical Name: N-(4-Diphenylphosphinophenyl)-N'-(3,6,9,12-tetraoxatridecyl)perylene-3,4,9,10-tetracarboxydiimide

Appearance: Dark red crystalline powder or solid
Purity: ≥90.0% (HPLC)
MW: 840.85, C51H41N2O8P

Storage Condition: 0-5ºC, protect from light
Shipping Condition:
ambient temperature

Product Description
Liperfluo, a perylene derivative containing oligooxyethylene, is designed and exclusively developed by Dojindo for a detection of lipid peroxides and emits intense fluorescence by a lipid peroxide specific oxidation in organic solvents such as ethanol. Among fluorescent probes that detect Reactive Oxigen Species(ROS), Liperfluo is the only compound that can specifically detect lipid peroxides. Since the excitation and emission wavelengths of the oxidized Liperfluo are 524 nm and 535 nm, respectively, both a photo-damage against a sample and an auto-fluorescence from the sample can be minimized. The tetraethyleneglycol group linked to one end of diisoquinoline ring helps its solubility and dispersibility to aqueous buffer. Though Liperfluo oxidized form is almost nonfluorescent in an aqueous media, it emits fluorescence in lipophilic sites such as in cell membranes. Therefore it can easily be applied
to lipid peroxide imaging by a fluorescence microscopy and a flow cytometric analysis for living cells.

                                Reaction of Liperfluo with lipid peroxide



Live cell imaging of lipid peroxide

  Procedure
1. Innocurate SH-SY5Y cells(6.0 x 105 cells/well) to a 6-well plate.
2. Incubate the plate at 37 ºC for overnight.
3. Add Liperfluo, DMSO solution(final conc. 20 μM) and incubate at
37 ºC for 15 min.
4. Add either Cumene Hydroperoxide(final conc. 100 μM) or AIPH*(final conc. 6 mM).
5. Incubate at 37 ºC for 2 hours.
6. Observe fluorescent by microscope**.

* AIPH: 2,2’-azobis-[2-(2-imidazolin-2-yl)propane]dihydrochloride
** Olympus IX-71 epifluorescent microscope, mirror unit: U-MNIBA3, exposure time: 10 sec, ISO: 800
   
Data was kindly provided from Dr. N. Noguchi, Doshisha University, System Life Science Laboratory.
K. Yamanaka and N. Noguchi et al., “A novel fluorescent probe with high sensitivity and selective detection of lipid hydroperoxides in cells”, RSC Advances, 2012, 2, 7894.



Flow cytometric analysis of lipid hydroperoxides in live cell

  Procedure
1. Innocurate SH-SY5Y cells(6.0 x 105 cells/well) to a 6-well plate.
2. Incubate the plate at 37 ºC for overnight.
3. Add Liperfluo, DMSO solution(final conc. 20 μM) and incubate at 37 ºC for 15 min.
4. Add either Cumene Hydroperoxide(final conc. 100 μM) or AIPH*(final conc. 6 mM).
5. Incubate at 37 ºC for 2 hours.
6. Wash cells with PBS.
7. Collect cells with PBA and analyse by flow cytometer**. 

* AIPH: 2,2’-azobis-[2-(2-imidazolin-2-yl)propane]dihydrochloride** BD FACSAriaTM I