The CD25
+
CD49d
-
Regulatory T Cell Isolation Kit was developed for the isolation of CD4
+
CD25
+
CD49d
-
regulatory T cells from human PBMCs in a two-step procedure.

Data and images for
CD25
+
CD49d
Regulatory T Cell Isolation Kit
, human

Figures

Figure 1

CD25
+
CD49d
regulatory T cells were isolated from human PBMCs using the CD25
+
CD49d
Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer.
PBMCs before separation
View details

Figure 1

CD25
+
CD49d
regulatory T cells were isolated from human PBMCs using the CD25
+
CD49d
Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer.
View details

Figure 1

CD25
+
CD49d
regulatory T cells were isolated from human PBMCs using the CD25
+
CD49d
Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer.
Pre-enriched CD25+CD49d- Treg cells
View details

Figure 1

CD25
+
CD49d
regulatory T cells were isolated from human PBMCs using the CD25
+
CD49d
Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer.
View details

Figure 1

CD25
+
CD49d
regulatory T cells were isolated from human PBMCs using the CD25
+
CD49d
Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer.
Isolated CD25+CD49d- Treg cells
View details

Figure 1

CD25
+
CD49d
regulatory T cells were isolated from human PBMCs using the CD25
+
CD49d
Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer.
View details

Figure 1

CD25
+
CD49d
regulatory T cells were isolated from human PBMCs using the CD25
+
CD49d
Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer.
Gated on CD4
+
cells.
View details

Figure 1

CD25
+
CD49d
regulatory T cells were isolated from human PBMCs using the CD25
+
CD49d
Regulatory T Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD4-FITC, CD25-PE, CD49d-Biotin and Anti-Biotin-APC, or CD4-FITC, CD25-PE, and Anti-FoxP3-APC and analyzed by flow cytometry using the MACSQuant® Analyzer.

Specifications for
CD25
+
CD49d
Regulatory T Cell Isolation Kit
, human

Overview

The CD25
+
CD49d
-
Regulatory T Cell Isolation Kit was developed for the isolation of CD4
+
CD25
+
CD49d
-
regulatory T cells from human PBMCs in a two-step procedure.

Detailed product information

Background information

CD49d is the α-chain of the integrin VLA-4 and is present on the majority of pro-inflammatory effector cells but absent on FoxP3
+
cells. Consequently, the depletion of CD49d
+
cells is a useful tool to isolate highly pure populations of CD25
+
CD49d
regulatory T cells.

Detailed separation procedure

The isolation of CD25
+
CD49d
regulatory T cells is performed in a two-step procedure. First, CD8
+
and CD49d
+
cells are magnetically labeled with a cocktail of CD8 and CD49d MicroBeads for depletion. In the second step, CD49d
regulatory T cells are labeled with CD25 MicroBeads and isolated by positive selection from the pre-enriched T cell fraction.
The depletion of CD49d
+
cells removes contaminating CD25
+
effector T cells
1
, thus providing highly pure populations of regulatory T cells.

Columns

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

Resources for
CD25
+
CD49d
Regulatory T Cell Isolation Kit
, human

Documents and Protocols

References for
CD25
+
CD49d
Regulatory T Cell Isolation Kit
, human

Publications

  1. Kleinewietfeld et al. (2009) CD49d provides access to "untouched" human Foxp3+ Treg free of contaminating effector cells. Blood 113: 827-836

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+
CD49d
Regulatory T Cell Isolation Kit
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