Miltenyi Biotec
CD8 MicroBeads, human
130-045-201
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The general aim of my research is to investigate the role of TCRs in the function of CTLs. For that we isolated TCR genes from T cell clones isolated from HIV patients and cloned them into lentiviral vectors to use for transduction of primary CD8 T cells isolated from naive patients. These transduced cells are then used for further study to evaluate their ability to recognize their cognate peptide and their killing capacity of infected cells. We selected this product because we had access to the AutoMACSpro, an automated machine used for cell separation compatible with these beads.
Separation of human CD8 positive cells from PBMCs
Human PBMCs
Count PBMCs, spin down. Add 80uL of running buffer and 20uL of microbeads per 10^7 PBMCs. Incubate at 4C for 15min. Wash the cells and resuspend in 3ml of running buffer. Load on to the LS column. Wash 3x with running buffer. Elute the cells using the plunger that comes with the LS column.
Could be possible to use less beads than the protocol recommends. If you are interested in saving some reagents titer the beads.
Good cell purity. High yield. Consistent results
None
Easy straight forward protocol with little optimization needed.
Expensive.
Great product to use for separation of CD8+ T cells from human PBMCs