MojoSort™ Human CD4 Nanobeads No Wash Protocol
Prepare cells from your tissue of interest without lysing erythrocytes .
In the final wash of your sample preparation , resuspend the cells in MojoSort™ Buffer by adding up to 4 mL in a 5 mL ( 12 x 75 mm ) polystyrene tube .
Note : Keep MojoSort™ Buffer on ice throughout the procedure .
Filter the cells with a 70 μm cell strainer , centrifuge at 300 x g for 5 minutes , and resuspend in an appropriate volume of MojoSort™ Buffer .
Count and adjust the cell concentration to 1 x 108 cells / mL .
Aliquot 100 μL of cell suspension ( 107 cells ) into a new tube .
Resuspend the beads by vortexing , maximum speed , 5 touches .
Add 10 μL of Nanobeads , mix well and incubate on ice for 15 minutes .
Scale up the volume accordingly if separating more cells ; for example , add 100 μL for 1 x 108 cells .
When working with less than 107 cells , use indicated volumes for 107 cells .
Resuspend the cells in 3 mL of MojoSort™ Buffer .
Optional : Take an aliquot before placing the tube in the magnet to monitor purity and yield .
Place the tube in the magnet for 5 minutes Pour out the liquid .
Resuspend labeled cells in appropriate buffer .
Repeat steps 6 – 8 on the labeled fraction 2 more times , for a total of 3 magnetic separations .
Optional : Take a small aliquot to monitor purity and yield .
If desired , pool the unlabeled fractions and process simultaneously with the positive labeled cells when assessing purity and yield .
