Protocol for Barcoding T Cells Combined with Timed Stimulations

Stimulation of naive T lymphocytes via the T cell receptor (TCR) induces distinct phosphorylation patterns that can be used to explore various signaling pathways within the cell. This protocol can be used to characterize different perturbations to the signaling pathways and the variations in time of...

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Detalles Bibliográficos
Autores principales: Kulhanek, Kayla R., Myers, Darienne R., Ksionda, Olga, Vercoulen, Yvonne, Romero-Moya, Damia, Roose, Jeroen P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580249/
https://www.ncbi.nlm.nih.gov/pubmed/33111105
http://dx.doi.org/10.1016/j.xpro.2020.100067
Descripción
Sumario:Stimulation of naive T lymphocytes via the T cell receptor (TCR) induces distinct phosphorylation patterns that can be used to explore various signaling pathways within the cell. This protocol can be used to characterize different perturbations to the signaling pathways and the variations in time of stimulation. Here, we provide a method of barcoding and consolidating a maximum of 24 different sample conditions using two florescent dyes. This single sample for phospho-staining and flow cytometry saves time and reagents. For complete details on the use and execution of this protocol, please refer to Krutzik and Nolan (2006), Krutzik et al. (2012), Vercoulen et al. (2017), Ksionda et al. (2018), and Myers et al. (2019).

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