A fast and sensitive method for isolation of detergent-resistant membranes from T cells.

TitleA fast and sensitive method for isolation of detergent-resistant membranes from T cells.
Publication TypeJournal Article
Year of Publication2005
JournalJournal of immunological methods
Volume305
Issue2
Pagination199-205
Date Published2005
ISSN0022-1759
Abstract

We describe a fast and sensitive method for isolation of detergent-resistant membranes (DRMs) from T cells by sucrose density gradient centrifugation using a smaller accumulated centrifugal force in a tabletop ultracentrifuge. Compared to previous reports, this method, which requires less biological material, is faster and permits quantitative separation of DRMs from other cellular membranes with good resolution. The method, which can be completed in 6 h, yields more than 80% of the total content of DRM-associated adaptor molecules LAT (linker for T cell activation), PAG/Cbp (protein associated with glycosphingolipid-enriched microdomains or Csk-binding protein) and LIME (Lck-interacting membrane protein) in low-density fractions using only 2x10(7) T cells.

URLhttp://linkinghub.elsevier.com/retrieve/pii/S0022-1759(05)00198-5
DOI10.1016/j.jim.2005.06.011
Short TitleJ Immunol Methods
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