Using EasySep™ Positive Selection Kits for Cell Depletion

The EasySep™ system allows you to use a positive selection kit to deplete the labeled cell type, by making changes to the separation scheme of the protocol.

Depletion Protocol:

  1. Prepare a single cell suspension according to the instructions in the kit-specific Product Information Sheet (PIS).
  2. Follow the instructions for reagent additions, incubation times and top-up volumes. Some titration of the reagent amounts added may be required.
  3. Place the tube containing the labeled cells into the magnet and set aside for 2x the amount of time stated in the PIS. For instance if the PIS indicates to “Place the tube (without lid) into the magnet and incubate at RT for 3 minutes”, extend this incubation to 6 minutes.
  1. Pick up the EasySep™ magnet and, in one continuous motion, invert the magnet and tube, pouring off the desired fraction into a new tube. The magnetically labeled unwanted cells will remain bound inside the tube, held by the magnetic field of the magnet. Hold the magnet and tube in inverted position for 2-3 seconds then, return to upright position. Do not shake or blot off any drops that may remain hanging from the mouth of the tube. Please refer to our Video: The EasySep™ Pour-off: A Simple and Effective Way to Isolate Cells to view the proper technique.
  2. Remove the original tube containing the magnetically labeled unwanted cells from the EasySep™ magnet and place the new tube, containing the desired enriched cells inside the magnet to perform a second round of magnetic separation. Repeat step 4*.
  3. The enriched cells are now ready for use.
*Optional: For some applications it may be desirable to perform only a single round of magnetic separation and stop the procedure after completion of Step 4. This will increase the overall cell recovery but may reduce the depletion efficiency of the targeted cells.

For the depletion of single or multiple unwanted cell types labeled with biotinylated antibodies, you may wish to consider our EasySep™ Mouse Streptavidin RapidSpheres™ Isolation Kit (Catalog # 19860).

For more information or assistance in optimizing your depletion protocol, please contact techsupport@stemcell.com.

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