MojoSort™ Streptavidin Nanobeads 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. 
Add appropriate amount of antibody or antibody cocktail to the cell suspension, mix well and incubate on ice for 15 minutes.
Note: The antibody volume to add should not exceed more than 20% of the 100 μL cell suspension volume.
Thus, for 100 μL of cell suspension do not add more than 20 μL of antibody.
If you need to add more than 20 μL of antibody, resuspend the cells in step 3 at a higher concentration.
For example, to add 50 μL of antibody, resuspend the cells to a final concentration of 2 x 108 cells/mL.
You can then aliquot 50 μL of cells and add 50 μL of antibody, mix well and incubate on ice for 15 minutes.
Always keep the total volume around 100 μL.
Add MojoSort™ Buffer up to 4 mL, centrifuge the cells at 300 x g for 5 minutes. 
Resuspend the cells in 100 μL of of MojoSort™ Buffer. 
Resuspend the beads by vortexing, maximum speed, 5 touches.
Add appropriate amount of pre-titrated Streptavidin Nanobeads, mix well and incubate on ice for 15 minutes.
Note: The Streptavidin Nanobeads volume to add should not exceed more than 20% of the 100 μL cell suspension volume.
Thus, for 100 μL of cell suspension do not add more than 20 μL of Nanobeads.
If you need to add more than 20 μL of Nanobeads, resuspend the cells in step 6 at a higher concentration.
For example, to add 50 μL of Nanobeads, resuspend the cells in 50 μL of MojoSort™ Buffer.
Always keep the total volume around 100 μL.
Add MojoSort™ Buffer up to 4 mL and centrifuge the cells at 300 x g for 5 minutes. 
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. 
Negative Selection: If you are interested in the untouched cells, collect the liquid in a new tube.
These are your cells of interest; do not discard.
If needed, repeat the magnetic separation on this cell fraction to increase the yield.
Positive Selection: If you are interested in the cells that are bound to the Nanobeads, pour off the liquid while the tube is in the magnet (negative fraction).
Then, remove the tube from the magnet and collect your cells.
Repeat steps 9 – 11 on the labeled fraction 2 more times, for a total of 3 magnetic separations to increase yield, if needed.
Optional: Take a small aliquot to monitor purity and yield.
Pool the unlabeled fractions and process simultaneously with the positive labeled cells when assessing purity and yield.
