The Double-negative T Cell Isolation Kit was developed for the isolation of double-negative T cells (CD4
CD8
CD56
CD3
+
TCRα/β
+
cells) from PBMCs by sequential sorting. The isolation is performed in a two-step procedure.

Data and images for Double-negative T Cell Isolation Kit, human

Figures

Figure 1

TCRα/β
+
CD4
CD8
double-negative T cells were isolated from human PBMCs by using the Double-negative T Cell Isolation Kit, one LD Column and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator. Cells were additionally fluorescently stained with Anti-Biotin-FITC.
PBMCs before separation
Pre-enriched double-negative T cells after depletion of CD4
+
, CD8
+
, and CD56
+
cells
View details

Figure 1

TCRα/β
+
CD4
CD8
double-negative T cells were isolated from human PBMCs by using the Double-negative T Cell Isolation Kit, one LD Column and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator. Cells were additionally fluorescently stained with Anti-Biotin-FITC.
View details

Figure 1

TCRα/β
+
CD4
CD8
double-negative T cells were isolated from human PBMCs by using the Double-negative T Cell Isolation Kit, one LD Column and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator. Cells were additionally fluorescently stained with Anti-Biotin-FITC.
Isolated double-negative T cells
View details

Figure 1

TCRα/β
+
CD4
CD8
double-negative T cells were isolated from human PBMCs by using the Double-negative T Cell Isolation Kit, one LD Column and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator. Cells were additionally fluorescently stained with Anti-Biotin-FITC.

Specifications for Double-negative T Cell Isolation Kit, human

Overview

The Double-negative T Cell Isolation Kit was developed for the isolation of double-negative T cells (CD4
CD8
CD56
CD3
+
TCRα/β
+
cells) from PBMCs by sequential sorting. The isolation is performed in a two-step procedure.

Detailed product information

Background information

The down-regulation of immune responses by regulatory T cells plays a key role in the induction of tolerance. Recently, it was reported that TCRα/β
+
CD4
CD8
double-negative T cells also have the ability to specifically down-regulate immune responses towards allo-antigens both in humans and mice. Double-negative T cells are found in lymphoid as well as in non-lymphoid tissues. They constitute about 1–3% of peripheral CD3
+
cells. It has been described that double-negative T cells can take up allo-MHC peptide complexes from antigen-presenting cells.

Detailed separation procedure

The isolation is performed in a two-step procedure. To remove all unwanted TCRα/β
+
cells, firstly the CD4
+
, CD8
+
, and CD56
+
cells are indirectly magnetically labeled and depleted. Labeling with Anti-TCRα/β-PE is performed simultaneously. In the second step, the flow-through fraction with the pre-enriched double-negative T cells is incubated with Anti-PE MicroBeads, and target cells are isolated by positive selection.

Columns

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

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