title: Western analysis of intracellular interleukin-8 in human mononuclear leukocytes.
abstract: Most cytokines are stored in the cytoplasm until their release into the extracellular environment; however, some cytokines have been reported to localize in the nucleus. Traditional whole cell extract preparation does not provide information about the intracellular localization of cytokines. Here, we describe how to prepare cytoplasmic and nuclear extracts that can be analyzed by immunoblotting. While in this chapter we use this method to analyze intracellular localization of interleukin-8 (IL-8) in human mononuclear leukocytes, this protocol is adaptable to any cell type or protein of interest.
authors.affiliation: Department of Anatomy and Structural Biology, Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Avenue, Bronx, NY, 10461, USA, veronika.miskolci@phd.einstein.yu.edu.
authors.forename: Veronika
authors.initials: V
authors.lastname: Miskolci
authors.forename: Louis
authors.initials: L
authors.lastname: Hodgson
authors.forename: Dianne
authors.initials: D
authors.lastname: Cox
authors.forename: Ivana
authors.initials: I
authors.lastname: Vancurova
mesh_terms: D015153:Blotting, Western; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D045424:Complex Mixtures / Q000737:chemistry; D003593:Cytoplasm / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005362:Ficoll; D015870:Gene Expression; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology
chemical_list: D045424:Complex Mixtures; D016209:Interleukin-8; D008070:Lipopolysaccharides; D005362:Ficoll