Isolation of intact RNA from murine CD4⁺ T cells after intracellular cytokine staining and fluorescence-activated cell sorting.

ABSTRACT: Intracellular cytokine staining (ICS) is a powerful method for identifying functionally distinct lymphocyte subsets, and for isolating these by fluorescence activated cell sorting (FACS). Although transcriptomic analysis of cells sorted on the basis of ICS has many potential applications, this is rarely performed because of the difficulty in isolating intact RNA from cells processed using standard fixation and permeabilization buffers for ICS. To address this issue, we compared three buffers shown previously to preserve RNA in nonhematopoietic cells subjected to intracellular staining for their effects on RNA isolated from T lymphocytes processed for ICS. Our results showed that buffers containing the recombinant ribonuclease inhibitor RNasin or high molar concentrations of salt yielded in

SUBMITTER: Kunnath-Velayudhan S 

PROVIDER: S-EPMC5878738 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature