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Cytokine Bioassays pp 285–293Cite as

Western Analysis of Intracellular Interleukin-8 in Human Mononuclear Leukocytes

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1172))

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.

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Acknowledgment

V.M. is supported by Cellular, Molecular Biology and Genetics training grant T32GM007491.

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Authors and Affiliations

  1. 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, Louis Hodgson & Dianne Cox

  2. Department of Biological Sciences, St. John’s University, Queens, NY, 11439, USA

    Ivana Vancurova

Authors
  1. Veronika Miskolci
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  2. Louis Hodgson
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  3. Dianne Cox
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  4. Ivana Vancurova
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Corresponding author

Correspondence to Veronika Miskolci .

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Editors and Affiliations

  1. Department of Biological Sciences, St. John's University, New York, New York, USA

    Ivana Vancurova

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Miskolci, V., Hodgson, L., Cox, D., Vancurova, I. (2014). Western Analysis of Intracellular Interleukin-8 in Human Mononuclear Leukocytes. In: Vancurova, I. (eds) Cytokine Bioassays. Methods in Molecular Biology, vol 1172. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0928-5_26

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  • DOI: https://doi.org/10.1007/978-1-4939-0928-5_26

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0927-8

  • Online ISBN: 978-1-4939-0928-5

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