Flow Cytometry Assay for Recycling of LFA-1 in T-lymphocytes
2018 (English)In: Bio-protocol, E-ISSN 2331-8325, Vol. 8, no 23, article id e3104Article in journal (Refereed) Published
Abstract [en]
To enable cells to move forward, cell surface integrins are internalized into an endosomal compartment and subsequently intracellularly transported to be re-exposed at a new site on the cell membrane. Leukocytes are the fastest migrating cell type in the human body, which express the leukocyte-specific integrin LFA-1. Here, we describe a flow cytometry-based assay that allows the quantification of LFA-1 internalization and its re-expression on the cell surface in T lymphocytes. An advantage of using flow cytometry-based assay over biochemical methods is the low number of needed cells. This protocol can be also used to measure recycling of other receptors.
Place, publisher, year, edition, pages
Bio-Protocol , 2018. Vol. 8, no 23, article id e3104
Keywords [en]
Flow cytometry, Receptor, LFA-1, Endocytosis, Exocytosis
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:oru:diva-72537DOI: 10.21769/BioProtoc.3104ISI: 000458028500009OAI: oai:DiVA.org:oru-72537DiVA, id: diva2:1289838
Funder
Swedish Research Council, K2010-80P-21592-01-4 K2010-80X-215917-01-4The Royal Swedish Academy of SciencesÅke Wiberg Foundation
Note
Funding Agencies:
Foundation Olle Engquist Byggmästare
I&A Lundberg Research Foundation
Royal Physiographic Society of Lund
Jeanssons Foundation
Kocks Foundation
P & U Schybergs Foundation
Gyllenstiernska Krapperup Foundation
Gustav V 80 Jubilee Fund
Österlund Foundation
Nanna Svartz and Crafoord awards
Anna-Greta Crafoord postdoctoral fellowship
2019-02-192019-02-192022-09-15Bibliographically approved