The CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II was developed for the isolation of CD4
+
CD25
+
CD127
dim/–
Treg cells from human PBMCs. The isolation is performed in a two-step procedure.

Data and images for
CD4
+
CD25
+
CD127
dim/-
Regulatory T Cell Isolation Kit II
, human

Figures

Figure 1

CD4
+
CD25
+
CD127
dim/–
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot.
PBMCs before separation
View details

Figure 1

CD4
+
CD25
+
CD127
dim/–
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot.
View details

Figure 1

CD4
+
CD25
+
CD127
dim/–
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot.
Pre-enriched CD4+CD127dim/- cells
View details

Figure 1

CD4
+
CD25
+
CD127
dim/–
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot.
Isolated CD4+CD25+CD127dim/- regulatory T cells
View details

Figure 1

CD4
+
CD25
+
CD127
dim/–
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot.
View details

Figure 1

CD4
+
CD25
+
CD127
dim/–
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot.
View details

Figure 1

CD4
+
CD25
+
CD127
dim/–
regulatory T cells were isolated from human PBMCs by using the CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II, an LD and two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD4-FITC, CD25-APC, and CD127-PE, or CD4-FITC, CD127-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression, except for the CD25-APC vs. CD4-FITC dot plot.

Specifications for
CD4
+
CD25
+
CD127
dim/-
Regulatory T Cell Isolation Kit II
, human

Overview

The CD4
+
CD25
+
CD127
dim/–
Regulatory T Cell Isolation Kit II was developed for the isolation of CD4
+
CD25
+
CD127
dim/–
Treg cells from human PBMCs. The isolation is performed in a two-step procedure.

Detailed product information

Background information

CD127, the α-chain of the IL-7 receptor, is expressed on the majority of mature T cells and plays an important role in their proliferation and differentiation. However, on regulatory T cells CD127 is absent and its expression inversely correlates with FoxP3 expression. Thus, CD127 can be used as an additional marker to discriminate between human regulatory and activated T cells.

Detailed separation procedure

First, the isolation of CD4
+
CD25
+
CD127
dim/–
regulatory T cells was performed with a cocktail of biotinylated antibodies and Anti-Biotin MicroBeads for the depletion of non-CD4
+
and CD127
high
cells. In the second step, the flow-through fraction of pre-enriched CD4
+
CD127
dim/–
T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4
+
CD25
+
CD127
dim/–
regulatory T cells.

Downstream applications

Isolation of CD4
+
CD25
+
CD127
dim/–
regulatory T cells from human PBMCs for further phenotypical or functional characterization.

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
+
CD127
dim/-
Regulatory T Cell Isolation Kit II
, human

References for
CD4
+
CD25
+
CD127
dim/-
Regulatory T Cell Isolation Kit II
, human

Publications

  1. Seddiki, N. et al. (2006) Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells. J. Exp. Med. 203: 1693-1700
  2. Liu, W. et al. (2006)
    CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4
    +
    T reg cells.
    J. Exp. Med. 203: 1701-1711
  3. Stockis et al. (2009) Membrane protein GARP is a receptor for latent TGF-beta on the surface of activated human Treg. Eur. J. Immunol. 39: 3315-3322

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CD4
+
CD25
+
CD127
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Regulatory T Cell Isolation Kit II
, human

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