Sensitive detection of GM1 lipid rafts and TCR partitioning in the T cell membrane

S. Thomas, R. S. Kumar, S. Casares, T. D. Brumeanu*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

18 Scopus citations


The cholesterol-rich lipid rafts on T cell membrane play important role in the formation of T cell receptor (TCR) signalosome upon receptor ligation. Analytical studies on the kinetics of lipid rafts formation and recruitment of protein receptors to lipid rafts are still limited by the use of a large number of cells. Herein, we describe a strategy for detecting fine alterations in the amount and distribution of glycosphingolipid (GM1) lipid rafts, and in the formation of GM1-TCR complexes in detergent-insoluble and -soluble compartments of the T cell membrane from a relative low number of cells. Using this strategy, we found that the GM1 moiety was physically associated with TCR in both detergent-insoluble and -soluble fractions. Shortly after ligation of CD3/TCR complex with a soluble CD3-ε mAb, the TCR was found mainly in the detergent-soluble fraction of the T cell membrane.

Original languageEnglish
Pages (from-to)161-168
Number of pages8
JournalJournal of Immunological Methods
Issue number1-2
StatePublished - 1 Apr 2003
Externally publishedYes


  • CD3-mediated TCR cross-linking
  • GM1 and TCR distribution
  • Glycosphingolipid GM1


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