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Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells
Previously published protocols for quantification of endosomal recycling are limited by the use of radioactive reagents, washing of cells in reducing buffers, or the requirement for large numbers of cells. Here, we describe a protocol for quantification of endosomal recycling using immunofluorescenc...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9038772/ https://www.ncbi.nlm.nih.gov/pubmed/35496808 http://dx.doi.org/10.1016/j.xpro.2022.101305 |
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author | Lonic, Ana Onglao, Winona Khew-Goodall, Yeesim |
author_facet | Lonic, Ana Onglao, Winona Khew-Goodall, Yeesim |
author_sort | Lonic, Ana |
collection | PubMed |
description | Previously published protocols for quantification of endosomal recycling are limited by the use of radioactive reagents, washing of cells in reducing buffers, or the requirement for large numbers of cells. Here, we describe a protocol for quantification of endosomal recycling using immunofluorescence that is optimized for EGFR in BT-549 breast cancer cells but could be applied to other RTKs and cell lines. Our protocol enables quick assessment of recycling and uses a relatively small number of cells. For complete details on the use and execution of this protocol, please refer to Lonic et al. (2021). |
format | Online Article Text |
id | pubmed-9038772 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-90387722022-04-27 Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells Lonic, Ana Onglao, Winona Khew-Goodall, Yeesim STAR Protoc Protocol Previously published protocols for quantification of endosomal recycling are limited by the use of radioactive reagents, washing of cells in reducing buffers, or the requirement for large numbers of cells. Here, we describe a protocol for quantification of endosomal recycling using immunofluorescence that is optimized for EGFR in BT-549 breast cancer cells but could be applied to other RTKs and cell lines. Our protocol enables quick assessment of recycling and uses a relatively small number of cells. For complete details on the use and execution of this protocol, please refer to Lonic et al. (2021). Elsevier 2022-04-13 /pmc/articles/PMC9038772/ /pubmed/35496808 http://dx.doi.org/10.1016/j.xpro.2022.101305 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Lonic, Ana Onglao, Winona Khew-Goodall, Yeesim Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells |
title | Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells |
title_full | Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells |
title_fullStr | Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells |
title_full_unstemmed | Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells |
title_short | Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells |
title_sort | quantifying egfr endosomal recycling via immunofluorescence in breast cancer cells |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9038772/ https://www.ncbi.nlm.nih.gov/pubmed/35496808 http://dx.doi.org/10.1016/j.xpro.2022.101305 |
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