Integrin Dependent RhoB Activation Assay Using Leukocytes

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 re...

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Bibliographic Details
Published inBio-protocol Vol. 8; no. 23; p. e3105
Main Authors Lehtonen, Janne, Potrzebowska, Katarzyna, Samuelsson, Malin, Svensson, Lena
Format Journal Article
LanguageEnglish
Published United States Bio-Protocol 05.12.2018
Bio-protocol LLC
Subjects
Biology
Leukocytes
LFA-1
Methods
Pull-down
RhoB
Rhoteikin
Small GTPases
Leukocytes
Pull-down
LFA-1
Rhoteikin
Small GTPases
RhoB
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Summary: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.
Bibliography:ObjectType-Article-1
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ISSN:2331-8325
2331-8325
DOI:10.21769/BioProtoc.3105