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Protocol for fluorescence-activated cell sorting of human EpCAM(+) lung cancer cells for gene expression analysis of Rac guanine-nucleotide exchange factors
Here, we describe a protocol for fluorescence-activated cell sorting (FACS) of human EpCAM(+) cells from fresh surgically resected specimens. We then use Q-PCR to identify specific molecular targets associated with the metastatic phenotype. This combined approach enables a qualitative and quantitati...
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/PMC9079335/ https://www.ncbi.nlm.nih.gov/pubmed/35542175 http://dx.doi.org/10.1016/j.xpro.2022.101367 |
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author | Sullivan, Neil T. Cooke, Mariana |
author_facet | Sullivan, Neil T. Cooke, Mariana |
author_sort | Sullivan, Neil T. |
collection | PubMed |
description | Here, we describe a protocol for fluorescence-activated cell sorting (FACS) of human EpCAM(+) cells from fresh surgically resected specimens. We then use Q-PCR to identify specific molecular targets associated with the metastatic phenotype. This combined approach enables a qualitative and quantitative gene expression analysis of lung cancer samples. We describe how to use the protocol for Rac GEFs, but it can be applied broadly to other molecular targets. For complete details on the use and execution of this protocol, please refer to Cooke et al. (2021) and Quatromoni et al. (2015). |
format | Online Article Text |
id | pubmed-9079335 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-90793352022-05-09 Protocol for fluorescence-activated cell sorting of human EpCAM(+) lung cancer cells for gene expression analysis of Rac guanine-nucleotide exchange factors Sullivan, Neil T. Cooke, Mariana STAR Protoc Protocol Here, we describe a protocol for fluorescence-activated cell sorting (FACS) of human EpCAM(+) cells from fresh surgically resected specimens. We then use Q-PCR to identify specific molecular targets associated with the metastatic phenotype. This combined approach enables a qualitative and quantitative gene expression analysis of lung cancer samples. We describe how to use the protocol for Rac GEFs, but it can be applied broadly to other molecular targets. For complete details on the use and execution of this protocol, please refer to Cooke et al. (2021) and Quatromoni et al. (2015). Elsevier 2022-05-02 /pmc/articles/PMC9079335/ /pubmed/35542175 http://dx.doi.org/10.1016/j.xpro.2022.101367 Text en © 2022 The Author(s) 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 Sullivan, Neil T. Cooke, Mariana Protocol for fluorescence-activated cell sorting of human EpCAM(+) lung cancer cells for gene expression analysis of Rac guanine-nucleotide exchange factors |
title | Protocol for fluorescence-activated cell sorting of human EpCAM(+) lung cancer cells for gene expression analysis of Rac guanine-nucleotide exchange factors |
title_full | Protocol for fluorescence-activated cell sorting of human EpCAM(+) lung cancer cells for gene expression analysis of Rac guanine-nucleotide exchange factors |
title_fullStr | Protocol for fluorescence-activated cell sorting of human EpCAM(+) lung cancer cells for gene expression analysis of Rac guanine-nucleotide exchange factors |
title_full_unstemmed | Protocol for fluorescence-activated cell sorting of human EpCAM(+) lung cancer cells for gene expression analysis of Rac guanine-nucleotide exchange factors |
title_short | Protocol for fluorescence-activated cell sorting of human EpCAM(+) lung cancer cells for gene expression analysis of Rac guanine-nucleotide exchange factors |
title_sort | protocol for fluorescence-activated cell sorting of human epcam(+) lung cancer cells for gene expression analysis of rac guanine-nucleotide exchange factors |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9079335/ https://www.ncbi.nlm.nih.gov/pubmed/35542175 http://dx.doi.org/10.1016/j.xpro.2022.101367 |
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