Equine Neutrophil Isolation

[MichelleT]

Hello,

I have been isolating neutrophils from human blood and my protocol works well. However, I have been finding the isolation much more difficult in equine blood. I am having trouble with the neutrophils clumping. I've searched the literature to find out what might be causing this, but I haven't found the answer. Does anyone have ideas?

[Zagami]

Neutrophils clumping can be observed in blood anticoagulated with citrate or heparin. Centrifuged sample at low speed and plasma replacement with PBS or saline at 37 °C, and as an alternative, holding blood specimen at 37 °C that prevented agglutination. This phenomenon can be linked presence of IgM on neutrophils surfaces.
A method for optimal and not contaminated neutrophils separation from the peripheral blood, from me experimented, you will find in:
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