IBA Lifesciences CD8+ CD62L+ CD45RO+ Tcm Fab Streptamer™ Isolation Kit MB

Description

• Strep-Tactin™ Magnetic Microbeads for Fab Streptamers™; 4 vials for 5 x 10⁸ cells each
• CD8 Fab-Strep, human for isolation of T-cells; 1 vial for 1 x 10⁹ cells
• CD62L Fab-Strep, human for isolation of T-cells; 1 vial for 5 x 10⁸ cells
• CD45RO Fab-Strep, human for isolation of T-cells; 1 vial for 2 x 10⁸ cells
• Biotin stock solution for Streptamer™ technology; 3 x 1 mL
• Buffer IS, Streptamer™ Washing Buffer, 10x concentrated; 2 x 50 mL
• Isolation of human CD8+ CD62L+ CD45RO+ central memory T cells is performed by serial triple positive selection.
• In a first step PBMCs are labeled with magnetic CD8 Fab Streptamers™.
• Labeled cells are isolated in a strong magnet (StrepMan, horizontal position) where they migrate toward the tube wall on the side of the magnet.
• This CD8 positive cell fraction is collected and cells are liberated from all labeling reagents by addition of biotin in a strong magnet (StrepMan, upright position).
• The magnetic Streptamers™ migrate toward the tube wall and the label-free cells remain in the supernatant.
• Biotin is removed by washing.The pure and label-free CD8 positive cells are subjected to a second positive selection with magnetic CD62L Fab Streptamers™ following the same procedure.
• The CD8+ CD62L+ T cells are then subjected to a third positive selection with CD45RO Fab Streptamers™.
• The resulting cell preparation is highly enriched with CD8+ CD62L+ CD45RO+ central memory T cells with a purity of more than 90%.
• No depletion steps and no columns are needed.