GE Healthcare Life Sciences
Ficoll-Paque Plus
17-1440-02
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We use this reagent to separate white blood cells from red blood cells using density. We routinely use this reagent to isolate leukocytes from human PMBC blood draws. In addition, it is gentle enough to isolate live cells from non-viable or unhealthy cells that can then be used for other applications, so we can work with the more viable and healthier cells.
Blood separation or enrichment of viable cells
Human PBMCs or T cell cultures
Everything must be moved slowly with very little perturbation to avoid mixing of gradients. We mix whole blood at a 1:1 ratio into PBS. Then layer 6 mL of blood mixture onto 3 mL of Ficoll. When pipetting ficoll, do not be too forceful. When layering blood onto the surface of the ficoll, pipette slowly enough such that the blood does not break into the ficoll layer. Carefully move tubes containing blood and ficoll into a centrifuge and spin at 400g for 30 min without any break to avoid disruption of gradient. Retrieve desired cell fraction.
Be gentle and work slow
We are able to isolate the leukocytes and we are also able to enrich for viable cells.
N/A
Easy enough to identify sample layers if done correctly
None
Great reliable way to manually isolate cells using a density gradient