Live cell immunogold labelling of RNA polymerase II

Labeling nuclear proteins with electron dense probes in living cells has been a major challenge due to their inability to penetrate into nuclei. We developed a lipid-based approach for delivering antibodies coupled to 0.8 nm ultrasmall gold particles into the nucleus to label RNA polymerase II. Focu...

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Detalles Bibliográficos
Autores principales: Orlov, Igor, Schertel, Andreas, Zuber, Guy, Klaholz, Bruno, Drillien, Robert, Weiss, Etienne, Schultz, Patrick, Spehner, Danièle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4321181/
https://www.ncbi.nlm.nih.gov/pubmed/25662860
http://dx.doi.org/10.1038/srep08324
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author Orlov, Igor
Schertel, Andreas
Zuber, Guy
Klaholz, Bruno
Drillien, Robert
Weiss, Etienne
Schultz, Patrick
Spehner, Danièle
author_facet Orlov, Igor
Schertel, Andreas
Zuber, Guy
Klaholz, Bruno
Drillien, Robert
Weiss, Etienne
Schultz, Patrick
Spehner, Danièle
author_sort Orlov, Igor
collection PubMed
description Labeling nuclear proteins with electron dense probes in living cells has been a major challenge due to their inability to penetrate into nuclei. We developed a lipid-based approach for delivering antibodies coupled to 0.8 nm ultrasmall gold particles into the nucleus to label RNA polymerase II. Focussed Ion Beam slicing coupled to Scanning Electron Microscopy (FIB/SEM) enabled visualization of entire cells with probe localization accuracy in the 10 nm range.
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spelling pubmed-43211812015-02-12 Live cell immunogold labelling of RNA polymerase II Orlov, Igor Schertel, Andreas Zuber, Guy Klaholz, Bruno Drillien, Robert Weiss, Etienne Schultz, Patrick Spehner, Danièle Sci Rep Article Labeling nuclear proteins with electron dense probes in living cells has been a major challenge due to their inability to penetrate into nuclei. We developed a lipid-based approach for delivering antibodies coupled to 0.8 nm ultrasmall gold particles into the nucleus to label RNA polymerase II. Focussed Ion Beam slicing coupled to Scanning Electron Microscopy (FIB/SEM) enabled visualization of entire cells with probe localization accuracy in the 10 nm range. Nature Publishing Group 2015-02-09 /pmc/articles/PMC4321181/ /pubmed/25662860 http://dx.doi.org/10.1038/srep08324 Text en Copyright © 2015, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Orlov, Igor
Schertel, Andreas
Zuber, Guy
Klaholz, Bruno
Drillien, Robert
Weiss, Etienne
Schultz, Patrick
Spehner, Danièle
Live cell immunogold labelling of RNA polymerase II
title Live cell immunogold labelling of RNA polymerase II
title_full Live cell immunogold labelling of RNA polymerase II
title_fullStr Live cell immunogold labelling of RNA polymerase II
title_full_unstemmed Live cell immunogold labelling of RNA polymerase II
title_short Live cell immunogold labelling of RNA polymerase II
title_sort live cell immunogold labelling of rna polymerase ii
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4321181/
https://www.ncbi.nlm.nih.gov/pubmed/25662860
http://dx.doi.org/10.1038/srep08324
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