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Measuring EGFR plasma membrane density, stability, internalization, and recycling in alive adherent cells by cell surface ELISA

EGFR cell surface density, stability, internalization, and recycling can be measured by cell surface ELISA (cs-ELISA). Performing this experiment on ice impedes receptor internalization; thus the physiological cell surface receptor levels can be measured by cs-ELISA. Cell surface EGFR levels are det...

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Detalles Bibliográficos
Autores principales: Kazan, Jalal M., Pollato-Blanco, Ariadna, Lukacs, Gergely L., Pause, Arnim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9213824/
https://www.ncbi.nlm.nih.gov/pubmed/35755125
http://dx.doi.org/10.1016/j.xpro.2022.101475
Descripción
Sumario:EGFR cell surface density, stability, internalization, and recycling can be measured by cell surface ELISA (cs-ELISA). Performing this experiment on ice impedes receptor internalization; thus the physiological cell surface receptor levels can be measured by cs-ELISA. Cell surface EGFR levels are detected by measuring Amplex Red fluorescence intensity. Although cell surface receptor levels can be measured by flow cytometry, cs-ELISA does not include cell dissociation steps that might affect cell surface receptor levels. For complete details on the use and execution of this protocol, please refer to Kazan et al. (2019).