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…

Offenlegungen

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

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Isolation of Lipids from Human Blood-Derived Neutrophils by Biphasic Separation. J. Vis. Exp. (Pending Publication), e21657, doi: (2023).

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