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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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
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author Kulhanek, Kayla R.
Myers, Darienne R.
Ksionda, Olga
Vercoulen, Yvonne
Romero-Moya, Damia
Roose, Jeroen P.
author_facet Kulhanek, Kayla R.
Myers, Darienne R.
Ksionda, Olga
Vercoulen, Yvonne
Romero-Moya, Damia
Roose, Jeroen P.
author_sort Kulhanek, Kayla R.
collection PubMed
description 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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spelling pubmed-75802492020-10-26 Protocol for Barcoding T Cells Combined with Timed Stimulations Kulhanek, Kayla R. Myers, Darienne R. Ksionda, Olga Vercoulen, Yvonne Romero-Moya, Damia Roose, Jeroen P. STAR Protoc Protocol 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). Elsevier 2020-07-22 /pmc/articles/PMC7580249/ /pubmed/33111105 http://dx.doi.org/10.1016/j.xpro.2020.100067 Text en © 2020. http://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
Kulhanek, Kayla R.
Myers, Darienne R.
Ksionda, Olga
Vercoulen, Yvonne
Romero-Moya, Damia
Roose, Jeroen P.
Protocol for Barcoding T Cells Combined with Timed Stimulations
title Protocol for Barcoding T Cells Combined with Timed Stimulations
title_full Protocol for Barcoding T Cells Combined with Timed Stimulations
title_fullStr Protocol for Barcoding T Cells Combined with Timed Stimulations
title_full_unstemmed Protocol for Barcoding T Cells Combined with Timed Stimulations
title_short Protocol for Barcoding T Cells Combined with Timed Stimulations
title_sort protocol for barcoding t cells combined with timed stimulations
topic Protocol
url 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
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