MojoSort™ Negative Selection Columns Protocol
Prepare a single cell suspension and resuspend the cells with ice cold cell separation buffer ( MojoSort™buffer recommended ) .
Pass the cells through a 70 μm filter , centrifuge ( 300 x g for 5 minutes ) , discard the supernatant and resuspend the cells in cell separation buffer .
Adjust the cell concentration to 1 x 108 cells / mL .
Aliquot 100 μL ( 107 cells ) into a new tube .
Add 10 μL of the pre - diluted Biotin - Antibody Cocktail , mix well and incubate on ice for 15 minutes .
Scale up the volume if separating more cells .
For example , add 100 μL of pre - diluted antibody cocktail for separating 1 x 108 cells in 1ml of buffer .
When working withless than 107 cells , use indicated volumes for 107 cells .
Add cell separation buffer up to 4 mL ; centrifuge the cells at 300 x g for 5 minutes .
Discard supernatant and resuspend in 100 μL of buffer .
Vortex the Streptavidin conjugated Nanobeads ( to resuspend ) at max speed , 5 touches , and prepare the dilutions to test .
Add 10 μL of pre - diluted Streptavidin Nanobeads .
Mix well and incubate on ice for 15 minutes .
Scale up the volume accordingly if separating more cells .
For example , add 100 μL of pre - diluted Nanobeads for separating 1 x 108 cells in 1ml of buffer .
When working with less than 107 cells , use indicated volumes for 107 cells .
Note : Depending on the isolation kit you are using , a wash step may be required here .
If you observe aggregates , filter the suspension .
To maximize yield , you can disrupt the aggregates by pipetting the solution up and down .
Resuspend the cells in appropriate amount of buffer .
At least 500 μL is needed for column separation , Note : There are several types of commercially available columns , depending on your application , choose the one that fits best your experiment .
See 'Guidelines' for choosing a column for your experiment .
