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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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Detalles Bibliográficos
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
Descripción
Sumario: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).