At what temperature, the centrifugation should be done to isolate PBMCs from whole blood using Ficol gradient centrifugation?
I found different protocols saying different temperatures during centrifugation step. So a bit confused.
Also please tell about the appropriate rcf and duration for centrifugation.
Thank you,
Centrifuge at 1200g for 10 min at RT for the buffy coat to form.
At 4C it is assumed that PBMCs may clump, thereby reducing the yield.
It is recommended that you use a swing bucket and centrifuge with brakes off, to minimize dispersal of the thin layer of the buffy coat. It takes a while for the centrifuge to stop after the completion of centrifugation cycle, though.
You may also use sepMate 50/ 15 from stem Cell technologies, wherein you can do your centrifugation with brakes on.
The temperature is always 4 oC.
Not sure what protocol you use, see below for a working one.
https://bio-protocol.org/e323
Thank you for your suggestions Sudeep Kumar Chinedu Anthony Anene
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