YILIMART - CD25 CD49d- Reg T Cell Iso Kit, human

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.

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.

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


Pre-enriched CD25 ⁺ CD49d ^- Treg cells

Isolated CD25 ⁺ CD49d ^- Treg cells

Gated on CD4 ⁺ cells.

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

Regulatory T cell research

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CD25 CD49d- Reg T Cell Iso Kit, human

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Background information

Detailed separation procedure

Columns

Figure 1

Selected references

Brochures and posters

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