Double-negative T Cell Isolation Kit, human

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Firstly, CD4+, CD8+, and CD56+ cells were labeled with the double-negative T cell biotin-antibody cocktail and anti-biotin MicroBeads. The magnetically labeled cells were separated using an LD Column on the autoMACS Separator. Secondly, the selected TCR (T cell receptor) α/β+CD4−CD8− cells present in the flow-through faction were incubated with Anti-PE MicroBeads and separated with MS column.	The double-negative T cells were retained within the column and eluted by removing the column from the magnetic field.

Protocol tips
Firstly, CD4+, CD8+, and CD56+ cells were labeled with the double-negative T cell biotin-antibody cocktail and anti-biotin MicroBeads. The magnetically labeled cells were separated using an LD Column on the autoMACS Separator. Secondly, the selected TCR (T cell receptor) α/β+CD4−CD8− cells present in the flow-through faction were incubated with Anti-PE MicroBeads and separated with MS column.
Downstream tips
The double-negative T cells were retained within the column and eluted by removing the column from the magnetic field.

Publication protocol

DNTs were isolated from the peripheral blood by using a double negative T cell isolation kit according to the manufacturer's instructions (130-092-614, Miltenyi, Germany). Firstly, CD4+, CD8+, and CD56+ cells were labeled with the double-negative T cell biotin-antibody cocktail and anti-biotin MicroBeads. The magnetically labeled cells were separated using an LD Column on the autoMACS Separator. Secondly, the selected TCR (T cell receptor) α/β+CD4−CD8− cells present in the flow-through faction were incubated with Anti-PE MicroBeads and separated with MS column. The double-negative T cells were retained within the column and eluted by removing the column from the magnetic field.

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