STEMCELL Technologies offers a wide range of EasySep™ negative selection kits for the isolation of untouched cells. While these kits are designed to obtain a balance of optimal purity and recovery, it is possible to adapt most EasySep™ kits' performance towards increased recovery of desired cells if required. Please note that since purity and recovery performance are inversely related, lower purity (the extent of which will be dependent on the cell type being enriched for) can be expected when adjusting the kit’s performance towards increased recovery.

To adjust the negative selection kit’s performance towards higher recovery, please execute the enrichment protocol (adding reagents, incubating and topping-up with buffer) as outlined in the kit-specific Product Information Sheet, with the following additional steps:

• After the magnetic separation, place the tube containing the poured-off enriched cell fraction aside.
• Remove the tube containing the magnetically-labeled unwanted cells from the magnet.
• Add recommended buffer, to the same volume used prior to the original separation.
• Thoroughly mix the sample to ensure that the cells are removed from the tube walls.
• Place the tube back into the magnet for another separation. Use the same incubation time recommended for the first separation.
• Pour off the supernatant into a new tube. This second pour-off should contain additional desired cells and can be combined with the enriched cells poured-off from the first separation.*

Note: If it is your first time attempting to adjust the enrichment for higher recovery, it is suggested that you do not combine both poured-off supernatant fractions until an aliquot from each has been assessed for desired cell purity via flow cytometry. This will help you assess if the second pour-off is of sufficient purity to be pooled with the first pour-off.

For further assistance and information, or to find out if a custom isolation product might be an option for your specific aim, please contact techsupport@stemcell.com.

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