Validation of a multicolor staining to monitor phosphoSTAT5 levels in regulatory T-cell subsets.

ABSTRACT:

Background

Regulatory T cells (Tregs) are key players in immune tolerance. They express the transcription factor FOXP3 and are dependent of the STAT5 signaling for their homeostasis. So far, the study of phosphorylated epitopes by flow cytometry required treating the cells with methanol, which is harmful for several epitopes.

Methods

Here we assessed whether the PerFix EXPOSE reagent kit (PFE)(Beckman Coulter) allowed monitoring the phosphorylation level of STAT5 in Treg subpopulations together with complex immunophenotyping. Results observed with the PFE kit were compared to those observed without cell permeabilization for surface markers, with paraformaldehyde permeabilization for non-phosphorylated intracellular epitopes, and with methanol-based permeabilization for phosp

SUBMITTER: Ehx G 

PROVIDER: S-EPMC4791230 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature