title: Detection of CXCR2 cytokine receptor surface expression using immunofluorescence.
abstract: The interleukin-8 (IL-8, CXCL8) chemokine, also known as the neutrophil chemotactic factor, is a cytokine that plays a key role in inflammatory response, cell proliferation, migration, and survival. IL-8 expression is increased not only in inflammatory disorders, but also in many types of cancer, including prostate cancer. IL-8 acts as a ligand for the C-X-C chemokine receptor 2 (CXCR2) protein present on the cell plasma membrane. Binding of the IL-8 ligand to the CXCR2 receptor results in an intracellular signaling pathway mediated by GTP binding proteins coupled to the receptor itself. Knowledge of the CXCR2 expression levels facilitates the understanding of the role and function of IL-8. In this chapter, we describe a protocol that uses the immunofluorescence method and confocal microscopy to analyze the CXCR2 surface expression in human prostate cancer cells. However, this protocol is easily adaptable to analyze the surface expression of other cytokine receptors in different cell types.
authors.affiliation: Department of Biological Sciences, St. John's University, 8000 Utopia Parkway, Queens, NY, 11439, USA.
authors.forename: Clarissa
authors.initials: C
authors.lastname: Lam
authors.forename: Mahmud Arif
authors.initials: MA
authors.lastname: Pavel
authors.forename: Parul
authors.initials: P
authors.lastname: Kashyap
authors.forename: Zahra
authors.initials: Z
authors.lastname: Salehi-Najafabadi
authors.forename: Victoria
authors.initials: V
authors.lastname: Valentino
authors.forename: Yong
authors.initials: Y
authors.lastname: Yu
mesh_terms: D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism*; D008297:Male; D018613:Microscopy, Confocal; D011467:Prostate / Q000378:metabolism* / Q000473:pathology; D011485:Protein Binding; D023063:Receptors, Interleukin-8B / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D016707:Tissue Fixation
chemical_list: D016209:Interleukin-8; D023063:Receptors, Interleukin-8B