Protocol for Barcoding T Cells Combined with Timed Stimulations

Kayla R Kulhanek; Darienne R Myers; Olga Ksionda; Yvonne Vercoulen; Damia Romero-Moya; Jeroen P Roose

doi:10.1016/j.xpro.2020.100067

Protocol for Barcoding T Cells Combined with Timed Stimulations

Kayla R Kulhanek, Darienne R Myers, Olga Ksionda, Yvonne Vercoulen, Damia Romero-Moya, Jeroen P Roose

Research output: Contribution to journal › Article › Academic › peer-review

6 Downloads (Pure)

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).

Original language	English
Article number	100067
Pages (from-to)	1-17
Journal	STAR protocols
Volume	1
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2020.100067
Publication status	Published - 18 Sept 2020

Access to Document

10.1016/j.xpro.2020.100067

1-s2.0-S266616672030054X-mainFinal published version, 2.75 MBLicence: CC BY-NC-ND

Cite this

@article{54f29ad758fc4ad2bfdfd0f7cba744b7,

title = "Protocol for Barcoding T Cells Combined with Timed Stimulations",

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).",

author = "Kulhanek, {Kayla R} and Myers, {Darienne R} and Olga Ksionda and Yvonne Vercoulen and Damia Romero-Moya and Roose, {Jeroen P}",

note = "Publisher Copyright: {\textcopyright} 2020",

year = "2020",

month = sep,

day = "18",

doi = "10.1016/j.xpro.2020.100067",

language = "English",

volume = "1",

pages = "1--17",

number = "2",

}

TY - JOUR

T1 - Protocol for Barcoding T Cells Combined with Timed Stimulations

AU - Kulhanek, Kayla R

AU - Myers, Darienne R

AU - Ksionda, Olga

AU - Vercoulen, Yvonne

AU - Romero-Moya, Damia

AU - Roose, Jeroen P

PY - 2020/9/18

Y1 - 2020/9/18

N2 - 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).

AB - 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).

U2 - 10.1016/j.xpro.2020.100067

DO - 10.1016/j.xpro.2020.100067

M3 - Article

C2 - 33111105

SN - 2666-1667

VL - 1

SP - 1

EP - 17

JO - STAR protocols

JF - STAR protocols

IS - 2

M1 - 100067

ER -

Protocol for Barcoding T Cells Combined with Timed Stimulations

Abstract

Access to Document

Fingerprint

Cite this