Protocol for Barcoding T Cells Combined with Timed Stimulations
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Updated: Dec 3, 2025
Spatial and Temporal Control of T Cell Activation Using a Photoactivatable Agonist
Published on: April 25, 2018
Kayla R Kulhanek1, Darienne R Myers1, Olga Ksionda1
1Department of Anatomy, University of California San Francisco (UCSF), 513 Parnassus Avenue, Room HSW-1326, San Francisco, CA 94143-0452, USA.
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).
