title: Analysis of the cell surface expression of cytokine receptors using the surface protein biotinylation method.
abstract: Cytokines are pleiotropic, low-molecular-weight proteins that regulate the immune responses to infection and inflammation. They stimulate the immune responses by binding to cytokine receptors on the cell plasma membrane. Thus, knowledge of the expression level of particular cytokine receptors on cell surface is crucial for understanding the cytokine function and regulation. One of the techniques to explore the membrane embedded cytokine receptors is cell surface biotinylation. Biotinylated surface proteins can be rapidly purified through the strong interaction between biotin and streptavidin. Here, we describe the procedure for surface biotinylation and purification of biotinylated cytokine receptors for further downstream analysis.
authors.affiliation: Department of Biological Sciences, St. John's University, 8000 Utopia Parkway, Queens, NY, 11439, USA.
authors.forename: Mahmud Arif
authors.initials: MA
authors.lastname: Pavel
authors.forename: Clarissa
authors.initials: C
authors.lastname: Lam
authors.forename: Parul
authors.initials: P
authors.lastname: Kashyap
authors.forename: Zahra
authors.initials: Z
authors.lastname: Salehi-Najafabadi
authors.forename: Gurpreet
authors.initials: G
authors.lastname: Singh
authors.forename: Yong
authors.initials: Y
authors.lastname: Yu
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry*; D019921:Biotinylation; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D011485:Protein Binding; D018121:Receptors, Cytokine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012685:Sepharose / Q000031:analogs & derivatives* / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D013388:Succinimides / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; C465543:N-hydroxysulfosuccimide; D018121:Receptors, Cytokine; D013388:Succinimides; C049538:streptavidin-agarose; D001710:Biotin; D012685:Sepharose