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An ESR, Mass Spectrometry and Fluorescence Microscopy Approach to Study the Stearic Acid Derivatives Anchoring in Cells |
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| Authors: | A. Grammenos M. Fillet M. Collodoro S. Lecart C. Quoilin M.-P. Fontaine-Aupart M. Hoebeke |
| Affiliation: | 1. Laboratory of Biomedical Spectroscopy, Department of Physics, B5, University of Liège, Sart-Tilman, 4000, Liège, Belgium 2. Laboratory of Analytical Pharmaceutical Chemistry, Department of Pharmacy, Drug Research Center, University of Liège, 1 av. de l’H?pital, 4000, Liège, Belgium 3. Laboratory of Histology-Cytology, Departement of Chemistry, University of Liège, Sart-Tilman, 4000, Liège, Belgium 4. Laboratory of Mass Spectrometry, GIGA-R and CART, Department of Chemistry, University of Liège, Sart-Tilman, 4000, Liège, Belgium 5. Center of Biomedical Photonic of LUMAT Federation, University of Paris-Sud, Orsay, France 6. Institut of Molecular Sciences of Orsay, University of Paris-Sud, Orsay, France |
| Abstract: | Lateral phase separations in biological membranes are of great interest, making electron spin resonance (ESR) spectroscopy combined with spin labeling a nondestructive and sensitive technique for the study of lipid rafts. It is currently accepted that the spin probe is localized on the plasma membrane. However, no study confirms this hypothesis. Herein, we report, for the first time, an accurate multispectral method for the quantification of the lipid spin label presence in every subcellular fraction. Cells were incubated with the 5-DOXYL stearic acid derivative and then subfractionated. Results of our multimodal spectroscopy approach ubiquitously demonstrate that the ESR spin label presence only sets in the plasma membranes. |
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