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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...

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Autores principales: Sullivan, Neil T., Cooke, Mariana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
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).
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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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