We already have the EasyStep magnet system from StemCell Technologies. We have used it for IP of specific receptor proteins using Thermofisher/Invitrogen Dynabeads and commercially available antibodies. I want to isolate Astrocytes from adult tissue (>P100) in specific brain regions for later RNAseq, and I'm interested in using the ACSA-2 kit (Miltenyi Biotec) if possible. I understand that for optimal yield, removal of myelin is essential, and Miltenyi also has a myelin removal kit. I'm loathe to spend another $2K on another magnet, (not to mention the $350 for just 25 columns). Do I have to use their specific columns and MACs stands with their beads or could I just use standard tubes and our StemCell magnet with their kits? With the dynabead system, the beads are released by a final wash step using a detergent (or in the case of IP for later western blot, by adding our loading buffer, which contains SDS). I am not entirely clear on how the LS or MS columns remove the beads from the positively-labeled cells? The video I watched suggested that the beads are small enough that they don't interfere with cell function or downstream applications and therefore do not need to be removed? They also argue that column-based sorting is superior to column-free. What is your experience? If you have done this before, or have specific recommendations as to how to proceed, your insights would be greatly appreciated.

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