protocol_307.txt

MojoSort™ Positive Selection Columns Protocol
Prepare a single cell suspension and resuspend the cells with ice cold cell separation buffer (MojoSort™buffer is 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.
Vortex the antibody-conjugated Nanobeads (to resuspend) at max speed, 5 touches, and prepare the dilutions to test.
Add 10 μL of pre-diluted conjugated 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 1 x 108 cells.
When working with less than 107 cells, use indicated volumes for 107 cells.
Note: Depending on the conjugated nanobead you are using, a wash step may be required here.
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.