Western analysis of intracellular interleukin-8 in human mononuclear leukocytes

Veronika Miskolci, Louis Hodgson, Dianne Cox, Ivana Vancurova

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)285-293
Number of pages9
JournalMethods in Molecular Biology
Volume1172
DOIs
StatePublished - 2014
Externally publishedYes

Keywords

  • Cytokines
  • Cytoplasmic extract
  • Immunoblotting
  • Interleukin-8
  • Intracellular localization
  • Nuclear extract

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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