Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9902292 | Journal of Immunological Methods | 2005 | 13 Pages |
Abstract
We produced the cardiotrophin-like cytokine/cytokine-like factor-1 complex tagged with a Bir A biotin ligase AviTag peptide substrate. This cytokine could be efficiently biotinylated in vitro with Bir A. It was subsequently validated as a sensitive tool for ciliary neurotrophic factor receptor detection by flow cytometry and for magnetic-activated cell sorting. It was also shown to allow the detection of a specific receptor by activated B cells. Whereas binding to cells expressing the ciliary neurotrophic factor receptor could be prevented by competition with ciliary neurotrophic factor, binding to B cells was not. The biotinylated cardiotrophin-like cytokine/cytokine-like factor-1 complex therefore represents a new reagent to study ciliary neurotrophic factor and cardiotrophin-like cytokine receptor expression and for the identification of the putative cardiotrophin-like cytokine B cell receptor. It further validates the use of biotin ligase catalysed biotinylation for the detection of cytokine receptors.
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Authors
Isabelle Cognet, Florence Guilhot, Mélanie Gabriac, Sylvie Chevalier, Younes Chouikh, Alexandra Herman-Bert, Angélique Guay-Giroux, Sophie Corneau, Giovanni Magistrelli, Greg C. Elson, Hugues Gascan, Jean-François Gauchat,