Integrin Dependent RhoB Activation Assay Using Leukocytes
2018 (English)In: Bio-protocol, E-ISSN 2331-8325, Vol. 8, no 23, article id e3105Article in journal (Refereed) Published
Abstract [en]
To be able to migrate, leukocyte needs to re-use its adhesion molecules to move forward. These adhesion molecules are called integrins and are intracellularly transported via endocytosis and exocytosis in order to translocate to a new site on the cell membrane. The intracellular transportation is regulated by different small GTPases including RhoB. Here we describe an activation assay of RhoB in leukocytes migrating on ICAM-1Fc coated dishes using commercially available Rhoteikin coated agarose beads. Although this is a specific protocol for LFA-1 induced RhoB activation, it can also be used for RhoB activation induced by other soluble and insoluble factors.
Place, publisher, year, edition, pages
Bio-Protocol , 2018. Vol. 8, no 23, article id e3105
Keywords [en]
Leukocytes, LFA-1, RhoB, Small GTPases, Rhoteikin, Pull-down
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-72536DOI: 10.21769/BioProtoc.3105ISI: 000458028500010OAI: oai:DiVA.org:oru-72536DiVA, id: diva2:1289845
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 Engvist Byggmästare
I & A Lundberg Research Foundation
Jeanssons Foundation
Kocks Foundation
P & U Schybergs Foundation
Gyllenstiernska Krapperup Foundation
Gustav V 80 Jubilee Fund
Österlund Foundation
Nanna Svartz, Crafoord Awards Anna-Greta Crafoord Foundation
Royal Physiographic Society of Lund
2019-02-192019-02-192022-09-15Bibliographically approved