The Naive Pan T Cell Isolation Kit was developed for the fast and efficient isolation of untouched naive T cells from human PBMCs.
Detailed separation procedure
For depletion of memory T cells and non-T cells, PBMCs are incubated with a cocktail of biotinylated CD45RO, CD14, CD15, CD16, CD19, CD25, CD34, CD36, CD57, CD123, anti-HLA-DR, CD235a (Glycophorin A), and CD244 antibodies. Optionally, biotinylated Anti-TCRγ/δ antibodies can be added for depletion of TCRγ/δ+ T cells. The cells are subsequently magnetically labeled with Anti-Biotin MicroBeads. CD61 MicroBeads are added for concurrent depletion of platelets.
Naive pan T cells isolated with the Naive Pan T Cell Isolation Kit can be used when direct labeling of cell surface molecules could interfere with downstream applications, e.g., in studies on naive T cell activation and in functional studies of DCs. Untouched naive pan T cells can also be used for further isolation of specific T cell subsets.
LS or autoMACS® Columns.
Separation of untouched naive T cells from PBMCs by depletion of non-T cells using the Naive Pan T Cell Isolation Kit, an LS Column, and a MidiMACS™ Separator. Cells were labeled with CD3-VioBlue, CD197-PE and CD45RA-FITC and analyzed by flow cytometry using the MACSQuant® Analyzer.
PBMCs before separation
Untouched naive T cells