Detection of Functionalized Sphingolipid Analogs in Detergent-Resistant Membranes of Immune Cells

Methods Mol Biol. 2021:2187:313-325. doi: 10.1007/978-1-0716-0814-2_18.

Abstract

The analysis of protein enrichment in the detergent-resistant membranes (DRMs) isolated from immune cells enables us to analyze a link between the membrane lipid dynamics and cell activation. Here, we describe the fractionation of detergent-resistant membranes and the correlative analysis of the enrichment of T cell receptor (TCR) and ω-azido-modified synthetic ceramide in those fractions upon TCR stimulation.

Keywords: Azido-modified ceramide; Detergent-resistant membranes (DRMs); Fluorescence analysis; Immunoblots; Sucrose gradient; Ultracentrifugation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Fractionation / methods
  • Cell Line, Tumor
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Ceramides / metabolism
  • Detergents / metabolism
  • Humans
  • Jurkat Cells
  • Membrane Lipids / metabolism
  • Membrane Proteins / metabolism
  • Receptors, Antigen, T-Cell / metabolism
  • Sphingolipids / metabolism*

Substances

  • Ceramides
  • Detergents
  • Membrane Lipids
  • Membrane Proteins
  • Receptors, Antigen, T-Cell
  • Sphingolipids