StemSep™ Reagents for Isolating Minor T Cell

Both major and minor T cell subsets can now be easily isolated by negative selection using StemSep™. With StemSep™, the unwanted cells are immunomagnetically labeled by a cocktail of patented crosslinking reagents called tetrameric antibody complexes. The unwanted cells are then efficiently eliminated from the cell suspension by passage through a magnetic separation column while the desired T cells are collected in the column effluent, highly enriched. The purified T cells have never been labeled with antibodies, thus avoiding undesirable side effects often associated with positive cell selection strategies such as blocking or activation.

Major T cell subsets (eg. CD3+, CD4+ or CD8+) are easily enriched using the appropriate standard StemSep™ T cell enrichment cocktail. StemCell Technologies also provides a range of individual tetrameric antibody complexes that target certain activation or development markers on T cells. Addition of one or more of these individual tetramers to the standard T cell enrichment cocktails allows the researcher to isolate many of the minor T cell subsets using a customized "Mix & Match" approach (see Tables 1 and 2).

The tables below show the range of human and murine T cell subsets that can currently be enriched with readily available StemSep™ reagents. In addition, StemCell Technologies can incorporate any murine IgG1 or biotinylated antibody from other sources into the StemSep™ system, allowing researchers to design their own unique cocktails.

Table 1. StemSepä Enrichment Cocktails for Human T Cell Subsets

Human T cell subset	Antibody Cocktails and Tetramers Required		Human T cell subset	Antibody Cocktails and Tetramers Required
	Standard Human Cocktail	Additional Tetramers		Standard Human Cocktail	Additional Tetramers
CD3+ T Cells	T Cell Enrichment	N/A	Memory CD4+	CD4+ T Cell Enrichment	CD45RA
CD4+ T Cells	CD4+ T Cell Enrichment	N/A	Resting CD4+	CD4+ T Cell Enrichment	one or more of:CD25CD69HLA-DR
CD8+ T Cells	CD8+ T Cell Enrichment	N/A	Naïve CD8+	CD8+ T Cell Enrichment	CD45RO
Naïve CD4+ T Cells	Naïve CD4+ T Cell Enrichment	N/A	Activated CD8+	CD8+ T Cell Enrichment	CD27
ab T Cells	T Cell Enrichment	TCR gd	Memory CD8+	CD8+ T Cell Enrichment	CD45RA
gd T Cells	T Cell Enrichment	TCR ab	Resting CD8+	CD8+ T Cell Enrichment	one or more of:CD25CD27CD69HLA-DR
Th1	CD4+ T Cell Enrichment	CD124 (IL-4R)

 

Table 2. StemSepä Enrichment Cocktails for Murine T Cell Subsets

Murine T cell subset	Antibody Cocktails and Tetramers Required		Murine T cell subset	Antibody Cocktails and Tetramers Required
	Standard Human Cocktail	Additional Tetramers		Standard Human Cocktail	Additional Tetramers
CD3+ T Cells	T Cell Enrichment	N/A	Memory CD4+	CD4+ T Cell Enrichment	DX5 (Pan NK)CD62L
CD4+ T Cells	CD4+ T Cell Enrichment	N/A	Resting CD4+	CD4+ T Cell Enrichment	CD25
CD8+ T Cells	CD8+ T Cell Enrichment	N/A	Naïve CD8+	CD8+ T Cell Enrichment	DX5 (Pan NK)CD44
DN Thymocytes	T Cell Enrichment	DX5 (Pan NK)CD4CD8CD25	Memory CD8+	CD8+ T Cell Enrichment	DX5 (Pan NK)CD62L
gd T Cells	T Cell Enrichment	DX5 (Pan NK)TCRab	Resting CD8+	CD8+ T Cell Enrichment	CD25
Naïve CD4+	CD4+ T Cell Enrichment	DX5 (Pan NK)CD44