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Biotinylation Assay to Determine LFA-1 Recycling in Motile T-Lymphocytes

Samuelsson, Malin LU and Svensson, Lena M. LU (2019) In Methods in molecular biology (Clifton, N.J.) 1930. p.115-122
Abstract

The cycles of internalization of the cell surface β2 integrin receptor lymphocyte function-associated antigen 1 (LFA-1) and its re-exposure on the plasma membrane are important for T-cell trafficking. Biotinylation of cells enables to measure surface expression of receptors, and after reducing surface biotin with reducing buffer, enables to measure the internalization of receptors. Here, by using biotin in combination with reducing buffer and recombinant intercellular adhesion molecule-1 (rICAM-1)-coated dishes and subsequent Western immunoblot analysis, we describe how to measure internalization of the LFA-1 receptor and its re-expression back to the cell surface in motile T-lymphocytes.

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author
and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Biotinylation, LFA-1 recycling, T-cell migration
in
Methods in molecular biology (Clifton, N.J.)
volume
1930
pages
8 pages
publisher
Springer
external identifiers
  • scopus:85059499042
  • pmid:30610605
ISSN
1940-6029
DOI
10.1007/978-1-4939-9036-8_14
language
English
LU publication?
yes
id
265d9edf-3736-4efe-b156-7d3e32aa3ec3
date added to LUP
2019-01-22 09:48:49
date last changed
2024-03-18 23:31:08
@article{265d9edf-3736-4efe-b156-7d3e32aa3ec3,
  abstract     = {{<p>The cycles of internalization of the cell surface β2 integrin receptor lymphocyte function-associated antigen 1 (LFA-1) and its re-exposure on the plasma membrane are important for T-cell trafficking. Biotinylation of cells enables to measure surface expression of receptors, and after reducing surface biotin with reducing buffer, enables to measure the internalization of receptors. Here, by using biotin in combination with reducing buffer and recombinant intercellular adhesion molecule-1 (rICAM-1)-coated dishes and subsequent Western immunoblot analysis, we describe how to measure internalization of the LFA-1 receptor and its re-expression back to the cell surface in motile T-lymphocytes.</p>}},
  author       = {{Samuelsson, Malin and Svensson, Lena M.}},
  issn         = {{1940-6029}},
  keywords     = {{Biotinylation; LFA-1 recycling; T-cell migration}},
  language     = {{eng}},
  pages        = {{115--122}},
  publisher    = {{Springer}},
  series       = {{Methods in molecular biology (Clifton, N.J.)}},
  title        = {{Biotinylation Assay to Determine LFA-1 Recycling in Motile T-Lymphocytes}},
  url          = {{http://dx.doi.org/10.1007/978-1-4939-9036-8_14}},
  doi          = {{10.1007/978-1-4939-9036-8_14}},
  volume       = {{1930}},
  year         = {{2019}},
}