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 .
