Hellow everyone
We try to isolate human eosinophils from fresh human blood samples using the new EasySep™ Human Eosinophil Enrichment Kit (Immunomagnetic negative selection) but end up with substantial platelets contamination!
The best we can get is 75% precent eosinophils and 25% pletlets even when we use human CD36 for additional platelets removal on the final kit cell pellets.
Does anyone have any suggestions or tips how can we achieve over 95% pure eosinophils?
We try to do several ex vivo activation experiments so any process that may lead to activation is problematic.
You should try low speed centrifugation (280-300g, 10 min, RT) before the EasySep. Most of the platelets should be in the upper layer (platelet-rich plasma-PRP-) while the cells will be in the pellet and buffy coat. Remove the yellowish upper layer (PRP, but don't touch/disturb the buffy coat) and replace with 1 or 2 volumes of PBS or other EasySep compatible buffer. Proceed with the EasySep.
Please let me know if it works,
best of luck,
Isabelle
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