Product Description
Liperfluo, a perylene derivative containing oligooxyethylene, is designed and exclusively developed by Dojindo for the detection of lipid peroxides. Liperfluo emits intense fluorescence by lipid peroxide specific oxidation in organic solvents such as ethanol. Among fluorescent probes that detect Reactive Oxygen 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, photo-damage and 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. Liperfluo's oxidized form is nearly nonfluorescent in an aqueous media and emits a strong 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. Liperfluo is used to monitor lipid peroxidation in ferroptosis research.
Reaction of Liperfluo with lipid peroxide
Properties of Liperfluo
Live cell imaging of lipid peroxide
|
Procedure
1. Innoculate 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 Eazobis-[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.
Flow cytometric analysis of lipid hydroperoxides in live cell
|
Procedure
1. Innoculate 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 Eazobis-[2-(2-imidazolin-2-yl)propane]dihydrochloride** BD FACSAriaTM I
|
Data was kindly provided from Dr. N. Noguchi, Doshisha University, System Life Science Laboratory.
|
Lipid peroxide of living cells
Cell line: L929
Microscope: Zeiss LSM510META
Filter type: FITC（GFP, Alexa488）wide filter
HFT UV/488
NFT490
BP505-550
|
Procedure:
- Prepare cell suspension (2.5 x 105 cell/well) in 35mm Glass bottom dish and incubate at 37 oC overnight in CO2.
- Discard the media and add new media containing Liperfluo (final conc. 1μM) .
- Incubate at 37 oC for 30 min in CO2.
- Discard the media add new media containing t-BHP (final conc. 250μM ).
- Incubate at 37 oC for 2 hours in CO2.
- Observe using confocal microscope.
Data was kindly provided from Dr. T. Kumagai and Dr. H. Imai, Kitasato University, School of Pharmacy.
Necrosis, apoptosis and autophagy is known as cell death-related processes. In 2012, Ferroptosis was proposed as one of new cell deaths. Ferroptosis is studied as non apoptotic cell death caused by accumulation of iron ion-dependent lipid peroxide. Liperfluo is used as a fluorescent prove which can detect intracellular lipid peroxide directly.
B. R. Stockwell et al., Cell, 2017, 171(2), 273.
V. E. Kagan et al., Nat. Chem. Biol., 2017, 13, (1), 81.
[Related Products]
1) N. Soh, T. Ariyoshi, T. Fukaminato, H. Nakajima, K. Nakano and T. Imato, "Swallow-tailed Perylene Derivative: a new Tool for Fluorescent Imaging of Lipid Hydroperoxides",
Org. Biomol. Chem.,
2007,
5, 3762.
2) K. Yamanaka, Y. Saito, J. Sakiyama, Y. Ohuchi, F. Oseto and N. Noguchi,
"A Novel Fluorescent Probe with High Sensitivity and Selective Detection of Lipid Hydroperoxides in Cells",
RSC Adv.,
2012, 2, (20), 7894.
3) V. E. Kagan, G. W. Mao, F. Qu, J. P. F. Angeli, S. Doll, C. S. Croix, H. H. Dar, B. Liu, V. A. Tyurin, V. B. Ritov, A. A. Kapralov, A. A. Amoscato, J. Jiang, T. Anthonymuthu, D. Mohammadyani, Q. Yang, B. Proneth, J. K. Seetharaman, S. Watkins, I. Bahar, J. Greenberger, R. K. Mallampalli, B. R. Stockwell, Y. Y. Tyurina, M. Conrad and H. Bayır,
"Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis",
Nature Chemical Biology.,
2017, 13, (1), 81.
4)Y. Nakashima, S. Ohta, A. M. Wolf,
"Blue light-induced oxidative stress in live skin.",
Free Radical Biology and Medicine.,
2017, 108, 300.
5) M. Tsugita, N. Morimoto and M. Nakayama,
"SiO2 and TiO2 nanoparticles synergistically trigger macrophage inflammatory responses",
Particle and Fibre Toxicology.,
2017, DOI 10.1186/s12989-017-0192-6.
6) K. Iuchi, A. Imoto, N. Kamimura, K. Nishimaki, H. Ichimiya, T. Yokota and S. Ohta,
"Molecular hydrogen regulates gene expression by modifying the free radical chain reactiondependent generation of oxidized phospholipid mediators",
Scientific Reports.,
2016, DOI: 10.1038/srep18971.
7) A. J. Clark, and H. R. Petty.,
"WO3/Pt nanoparticles promote light-induced lipid peroxidation and lysosomal instability within tumor cells.", Nanotechnology., 2016, 27, (7), 075103.
8) T. Otani, M. Matsuda, A. Mizokami, N. Kitagawa, H. Takeuchi, E. Jimi, T. Inai and M. Hirata,
"Osteocalcin triggers Fas/FasL-mediated necroptosis in adipocytes via activation of p300",
Cell Death Dis.,
2018, 9, 1194.
9) H.H. Dar, Y.Y. Tyurina, K. Mikulska-Ruminska, I. Shrivastava, H.C. Ting, V.A. Tyurin, J. Krieger, C.M. St Croix, S. Watkins, E. Bayir, G. Mao, C. Ambruster, A. Kapralov, H. Wang, M.H. Parsek, T.S. Anthonymuthu, A.F. Ogunsola, B.A. Flitter, C.J. Freedman, J.R. Gaston, T.R. Holman, J.M. Pilewski, J.S. Greenberger, R.K. Mallampalli, Y. Doi, J.S. Lee, I. Bahar, J.M. Bomberger, H. Bayır, V.E. Kagan,
"Pseudomonas aeruginosa utilizes host polyunsaturated phosphatidylethanolamines to trigger theft-ferroptosis in bronchial epithelium.",
J. Clin. Invest..,
2018,DOI: 10.1172/JCI99490. .
10) H. Alborzinia, T. I. Ignashkova, F. R. Dejure, M. Gendarme, J. Theobald, S. Wolfl, R. K. Lindemann and J. H. Reiling ,
"Golgi stress mediates redox imbalance and ferroptosis in human cells",
Commun Biol..,
2018,
1, (210), DOI: 10.1038/s42003-018-0212-6.
11) W. Wang, M. Green, J. E. Choi, M. Gijon, P. D. Kennedy, J. K. Johnson, P. Liao, X. Lang, I. Kryczek, A. Sell, H. Xia, J. Zhou, G. Li, J. Li, W. Li, S. Wei, L. Vatan, H. Zhang, W. Szeliga , W. Gu, R. Liu, T. S. Lawrence, C. Lamb, Y. Tanno, M. Cieslik, E. Stone, G. Georgiou, T. A. Chan, A. Chinnaiyan, W. Zou,
"CD8+ T cells regulate tumour ferroptosis during cancer immunotherapy.",
Nature.,
2019,
569(7755), 270-274.
12) M. Gao, J. Deng, F. Liu, A. Fan, Y. Wang, H. Wu, D. Ding, D. Kong, Z. Wang, D. Peer, Y. Zhao,
'Triggered ferroptotic polymer micelles for reversing multidrug resistance to chemotherapy',
Biomaterials.,
2019,
233, 119486.
13) N. Wang, GZ. Zeng, JL. Yin, ZX. Bian,
Artesunate activates the ATF4-CHOP-CHAC1 pathway and affects ferroptosis in Burkitt's Lymphoma',
Biochem. Biophys. Res. Commun.,
2019,
519, (3), 533-539.
14) T.Tsukui, Takayuki Tsukui, Z. Chen, H. Fuda, T. Furukawa, K. Oura, T. Sakurai, S. Hui, H. Chiba,
'Novel Fluorescence-Based Method To Characterize the Antioxidative Effects of Food Metabolites on Lipid Droplets in Cultured Hepatocytes',
J. Agric. Food Chem.,
2019,
67, (35), 9934-9941.
15) P. K. Mishra, A. Adameova, J. A. Hill, C. P. Baines, P. M. Kang, J. M. Downey, J. Narula, M. Takahashi, A. Abbate, H. C. Piristine, S. Kar, S. Su, J. K. Higa, N. K. Kawasaki and T. Matsui,
Guidelines for evaluating myocardial cell death',
Am. J. Physiol.: Heart Circ. Physiol. .,
2019,
317, (5), H891-H922 .
16) C. Liang, X. Zhang, M. Yang and X. Dong,
Recent Progress in Ferroptosis Inducers for Cancer Therapy",
Adv. Mater..,
2019,doi.org/10.1002/adma.201904197.
17) B. R. Stockwell and X. Jiang,
The Chemistry and Biology of Ferroptosis",
Cell Chem. Biol..,
2020,
27.
18) O. Beharier, V. A. Tyurin, J. P. Goff, J. Guerrero-Santoro, K. Kajiwara, T. Chu, Y. Y. Tyurina, C. M. St Croix, C. T. Wallace, S. Parry, W. T. Parks, V. E. Kagan, and Y. Sadovsky,
"PLA2G6 guards placental trophoblasts against ferroptotic injury",
Proc. Natl. Acad. Sci. U.S.A.,
2020, doi.org/10.1073/pnas.2009201117.
19) P. P. Yee, Y. Wei, Soo-Yeon Kim, T. Lu, S. Y. Chih, C. Lawson, M. Tang, Z. Liu, B. Anderson, K. Thamburaj, M. M. Young, D. G. Aregawi, M. J. Glantz, B. E. Zacharia, C. S. Specht, Hong-Gang Wang and W. Li,
"Neutrophil-induced ferroptosis promotes tumor necrosis in glioblastoma progression",
Nat. Commun.,
2020, doi.org/10.1038/s41467-020-19193-y.