Biolegend
MojoSort™ Streptavidin Nanobeads
480016
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We use a mouse model to study CD4 and CD8 T cell signaling and activation. Pure CD4 and CD8 are needed for RT-PCR, Western blot, and all other functional assays. So it is necessary to perform the CD4 and CD8 purification. We are currently using this product and get very good purity and yield.
Cell Isolation/Separation
Splenocytes
Biotin antibody cocktail
MojoSort Buffer
Streptavidin Nanobeads
NA
Unbound CD4 or CD8 purity is usually 95%.
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The purification steps are : 1. Resuspend the cells in MojoSort™ Buffer in a 5mL (12 x 75 mm) polypropylene tube, 107 cells/100µL. 2. Add 10l of antibody cocktail to 100l cell suspension, incubate on ice for 15 minutes. 3. Resuspend Streptavidin Nanobeads by vortexing. Add 10l beads to the tube, mix well, and incubate on ice for 15 minutes. 4. Add 2ml MojoSort™ Buffer to the tube and mix well. 5. Place the tube in the magnet for 5 minutes. 6. Collect the liquid in a new tube while the tube is in the magnet. The cells in the liquid are untouched cells (negative selection). If you are interested in the cells that are bound to the Nanobeads (positive selection), remove the tube from the magnet and collect the cells.
Good purity and reasonable yield
Easy method and reproducible