Analysis of Nucleocytoplasmic Protein Shuttling by Imaging Flow Cytometry

Fasler-Kan, Elizaveta; Baiken, Yeldar; Vorobjev, Ivan A.; Barteneva, Natasha S. (2015). Analysis of Nucleocytoplasmic Protein Shuttling by Imaging Flow Cytometry. In: Barteneva, Natasha S.; Vorobjev, Ivan A. (eds.) Imaging Flow Cytometry - Methods and Protocols. Methods in Molecular Biology: Vol. 1389 (pp. 127-137). New York: Springer Science + Business Media (Humana Press) 10.1007/978-1-4939-3302-0_8

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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

Item Type:

Book Section (Book Chapter)

Division/Institute:

04 Faculty of Medicine > Department of Gynaecology, Paediatrics and Endocrinology (DFKE) > Clinic of Paediatric Surgery

UniBE Contributor:

Fasler-Kan, Elizaveta

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1064-3745

ISBN:

978-1-4939-3300-6

Series:

Methods in Molecular Biology

Publisher:

Springer Science + Business Media (Humana Press)

Language:

English

Submitter:

Christoph Steffen

Date Deposited:

15 Feb 2016 09:50

Last Modified:

15 Feb 2017 13:38

Publisher DOI:

10.1007/978-1-4939-3302-0_8

Uncontrolled Keywords:

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

URI:

https://boris.unibe.ch/id/eprint/76188

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