Use this kit to isolate highly pure CD4+ CD25+ regulatory T-cells (Treg) that express the intracellular transcription factor Foxp3. The CD4+ CD25- cell fraction can be used as effector cells in downstream inhibitory assays. You can expect >95% purity (CD4+ CD25+ expression) and >80% Foxp3 expression.

Isolate bead- and antibody-free human CD4+ CD25+ Foxp3+ Treg cells
• Treg cells can be expanded with Dynabeads™ CD3/CD28 Treg Expander and remain
Foxp3+.
• Treg cells can be used in any downstream applications (e.g., suppression assays)

Methodology
The isolation procedure includes the following steps:
• Negative isolation of untouched human CD4+ T-cells from PBMC.
• Positive isolation of CD25+ T-cells from the untouched CD4+ T-cell subset.
• Removal of beads from the CD4+CD25+ Treg cells with DETACHaBEAD™

Starting sample
PBMC