We expect collect 10 ml of peripheral blood in order to isolate plasma and PBMC , after a density gradient separation with Lymphoprep using sep mate tubes. The collected plasma and PBMC will be used to study the proteins. Which additive would be appropriate for this application? EDTA, Sodium Citrate or Heparin.
unsure of your utility of isolations,
I used to follow,
EDTA : for exvivo flowcytometry staining ( cell and nuclear)
Heparin: for isolation of PBMCs and stimulations, as well as whole blood stimulation with antigens
Hi, I always use EDTA, but my target is RNA. In my approaches Heparin is an evil.
They should all work for anticoagulation , but check the endotoxin level with heparin, if you intend to use the isolated PBMC for activation studies. I've successfully used SODIUM-heparin vacuettes for years for cell/plasma isolation for immunological assays, but this may interfere with subsequent PCR reactions. As EDTA chelates key ions, this may effect subsequent cellular and enzymic reactions in assays at high plasma concentrations. While sodium citrate collection is essential for cell isolation for subsequent cellular assays with parasites eg plasmodium falciparum.
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