Analysis of nucleocytoplasmic protein shuttling by imaging flow cytometry

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Many intracellular signal transduction events involve the reversible shuttling of proteins between the cytoplasm and the nucleus. Study of these processes requires imaging information on the protein localization in a given cell and a large number of measurements to obtain sufficient statistics on the protein localization in the whole population. The protocol describes method for quantitative imaging flow cytometry analysis of intracellular distribution of NF-kappaB in ARPE-19 cells stained with specific fluorochrome-conjugated antibodies. The described technique alone or in combination with standard flow cytometry methods can be applied to study any protein undergoing translocation from cytoplasm into the nucleus in a variety of cell lines as well as in heterogeneous primary cell populations.

Original languageEnglish
Pages (from-to)127-137
Number of pages11
JournalMethods in molecular biology (Clifton, N.J.)
Volume1389
DOIs
Publication statusPublished - 2016

Keywords

  • ARPE-19 cell line
  • Flow cytometry
  • Imaging cytometry
  • NF-kappaB
  • Nucleocytoplasmic shuttling
  • Nucleocytoplasmic translocation
  • References
  • Retinal pigment cells

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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