[Histonet] Collecting PBMCs

From:annamaria touvra

Goodmorning everyone,
  I would like some help concerning the collection/separation of Peripheral Mononuclear Cells (PBMCs) from whole blood.
  If anyone has done it with HIstopaque-1077 of Sigma-Aldrich, I would like to know which is the appropriate ratio of Histopaque and whole blood, at what temperature is the centrifugation done, how many rpm are used and for how long. The procedure mentions to use 15ml centrifuge tubes. What will happen if I'll use tubes of 5 or 10 ml?
  If there is anything else making the separation better I would appreciate any comment.
  Thank you in advance,
  Have a nice week full of joy in work,

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