The CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit was developed for the isolation of CD45RA
+
regulatory T cells (Treg) from human PBMCs in a two-step procedure.

Data and images for
CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit
, human

Figures

Figure 1

CD4
+
CD25
+
CD45RA
+
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit, an LD, and two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. The isolated cells were fluorescently stained with CD4, CD25, and either CD45RA or Anti-FoxP3 antibodies. Dot plots show data gated on viable lymphocytes (C) or viable CD4
+
lymphocytes (A, B).
Viable CD4+ lymphocytes
View details

Figure 1

CD4
+
CD25
+
CD45RA
+
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit, an LD, and two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. The isolated cells were fluorescently stained with CD4, CD25, and either CD45RA or Anti-FoxP3 antibodies. Dot plots show data gated on viable lymphocytes (C) or viable CD4
+
lymphocytes (A, B).
View details

Figure 1

CD4
+
CD25
+
CD45RA
+
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit, an LD, and two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. The isolated cells were fluorescently stained with CD4, CD25, and either CD45RA or Anti-FoxP3 antibodies. Dot plots show data gated on viable lymphocytes (C) or viable CD4
+
lymphocytes (A, B).
Viable lymphocytes
View details

Figure 1

CD4
+
CD25
+
CD45RA
+
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit, an LD, and two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. The isolated cells were fluorescently stained with CD4, CD25, and either CD45RA or Anti-FoxP3 antibodies. Dot plots show data gated on viable lymphocytes (C) or viable CD4
+
lymphocytes (A, B).

Specifications for
CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit
, human

Overview

The CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit was developed for the isolation of CD45RA
+
regulatory T cells (Treg) from human PBMCs in a two-step procedure.

Detailed product information

Background information

Regulatory CD4
+
CD25
+
T cells isolated from the CD45RA
+
naive T cell compartment were shown to be optimal for
in vitro
expansion. These expanded Treg cells maintained the FoxP3
+
phenotype and their suppressive activity, whereas CD45RA
regulatory T cells from the memory T cell compartment lost FoxP3 expression and showed only modest suppressive activity.
1

Detailed separation procedure

The isolation is performed in a two-step procedure. First, non-CD4
+
and memory T cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted over a MACS
®
Column. In the second step, the flow-through fraction of pre-enriched CD4
+
CD45RA
+
T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4
+
CD25
+
CD45RA
+
T cells.

Downstream applications

CD4
+
CD25
+
CD45RA
+
regulatory T cells can be isolated from human PBMCs with this kit, for example, for further phenotypical or functional characterization and expansion.

Columns

For the first magnetic separation (depletion): LD or autoMACS
®
Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

Resources for
CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit
, human

Documents and Protocols

References for
CD4
+
CD25
+
CD45RA
+
Regulatory T Cell Isolation Kit
, human

Publications

  1. Hoffmann, P. et al. (2006)
    Only the CD45RA
    +
    subpopulation of CD4
    +
    CD25
    high
    T cells gives rise to homogeneous regulatory T-cell lines upon
    in vitro
    expansion.
    Blood 108: 4260-4267

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CD4
+
CD25
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, human

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