Isolation of Lipids from Human Blood-Derived Neutrophils by Biphasic Separation
Published: October 31, 2023
Abstract
Source: Brogden, G., et al. Methods to Study Lipid Alterations in Neutrophils and the Subsequent Formation of Neutrophil Extracellular Traps. J. Vis. Exp. (2017)
This video demonstrates lipid isolation from human neutrophils by a biphasic solvent system using methanol and chloroform.
Protocol
1. Lipid Isolation and Analysis of Human Blood-derived Neutrophils Take neutrophils and place them on ice. On the ice, pipette the neutrophils (present in methanol and chloroform solution) to a 15 mL screw-cap glass tube with a polytetrafluoroethylene (PTFE) seal and homogenize them by shaking for 1 min. Use glass tubes to prevent the lipids from binding to plastic surfaces. Use PTFE caps to prevent contamination from rubber/plastic. Add 2 mL of methanol followed 1 min lat…
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The authors have nothing to disclose.
Materials
| 10 µl syringe | Hamilton | 701 NR 10 µl | |
| Cannula 26G | Braun | 4657683 | |
| Chloroform | Carl Roth | 7331.1 | |
| Methanol | Carl Roth | 7342.1 | |
| Water | Carl Roth | 3255.1 | Endotoxin-free |
