Don't Want to Use Serum? Alternatives for Cell Separation Buffers

When isolating cells using EasySep™ or RoboSep™ cell separation systems, the traditionally recommended buffer is 2% FBS in Phosphate Buffered Saline (PBS, Ca2+ and Mg2+ free), with 1 mM EDTA. For your convenience, you can purchase this buffer ready-made from STEMCELL Technologies (EasySep™ Buffer, 1 L, Catalog #20144 or RoboSep™ Buffer, 250 mL, Catalog #20104).

However, if you prefer to avoid the use of serum, you could use an alternative buffer consisting of PBS (Ca2+ and Mg2+free), supplemented with 0.5% BSA or HSA, and 2 mM EDTA. We provide a serum-free buffer which contains 0.5% BSA (RoboSep Buffer 2, 250 mL, Catalog #20164). If required, you could also substitute the PBS for Hank's Balanced Salt Solution (HBSS, Ca2+ and Mg2+ free, Catalog #37250).

We cannot recommend using other media (such as RPMI, DMEM/F12 or DMEM) as the buffer for cell separations, as we have not tested this. Our cell separation kits have only been validated using the recommended buffers above. More importantly, most media (e.g. DMEM) is not buffered, unless it's maintained in an environment with 5-10% CO2. Outside of a CO2 incubator, the pH of the media will likely change, which is not optimal.

For further assistance and information, please contact techsupport@stemcell.com.