Protocol for Barcoding T Cells Combined with Timed Stimulations
Publication date
2020-09-18
Authors
Kulhanek, Kayla R
Myers, Darienne R
Ksionda, Olga
Vercoulen, Yvonne
Romero-Moya, Damia
Roose, Jeroen P
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cc_by_nc_nd
Abstract
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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Journal Article
Citation
Kulhanek, K R, Myers, D R, Ksionda, O, Vercoulen, Y, Romero-Moya, D & Roose, J P 2020, 'Protocol for Barcoding T Cells Combined with Timed Stimulations', STAR protocols, vol. 1, no. 2, 100067, pp. 1-17. https://doi.org/10.1016/j.xpro.2020.100067