We currently use a standard PBMC protocol, first centrifuging the citrate blood tubes and collecting plasma, followed by laying the remaining blood components diluted with HBSS over lymphoprep. This protocol has worked well for us, however, we were hoping to save time by trying the Sepmate tubes; we still would like to remove plasma before using the sepmate tubes, to keep the plasma data comparable with past studies. Has anyone done this before?
Hi Destanie--you can absolutely do this. In order for this protocol to work, you will need to replace the removed plasma with PBS. For example:
Starting material is 20 mL of whole blood
Spin whole blood to generate a buffy coat
Remove plasma (say 7 mL)
Replace removed plasma with PBS (7 mL)
Dilute blood with equal volume with PBS as you would before any density spin (add 20 more mL of PBS)
Proceed with SepMate protocol as usual
I hope this helps. If you have more questions find me on LinkedIn!