Hello,
I am planning to perform a cytotoxic assay with porcine splenocytes as effector cells. I have tried to find the proper concentration of porcine IL-2 that I should use for NKs stimulation but I think there is not a consensus for that. Does anyone have experience with this type of experiments? In this case, what concentration of porcine IL-2 you normally use?
Thanks in advance.
Iker
In general, you will aim to activate your cells with 100-200 IU/mL of IL-2. However, every experiment has its own set-up depending on the cell type and your target. Try to do different concentrations and to optimize your protocol based on the best results. Good luck!!
Dear Rehab,
Thank you for your answer. Actually, I have used IL-2 at 100 IU/mL. Once I have analysed the results I will know whether I have to try different concentration of IL-2.
Best wishes
Iker
Activation for killing assay require 3-5 days in 50-100 units/mL of IL-2. for other downstream experiments you optimise for the best outcome. you may have to titrate your IL-2 effect.
Also it depends on the manufacturer's supplier and source of recombinant IL-2 patia pastors or Ecoli
for example IL-2 from R&D work fine at 100 Units. we struggled with other manufacturer to get the same effect even using their metrics.
remember that fast sever activation will make your Nk cells go to apoptosis faster.
Dear Yasser,
Thank you very much for your answer. I really appreciate your help with this.
Best wishes
Iker
Dear Iker,
At iQ Biosciences for all of our ADCC assays for drug therapeutic screening, we use clinical grade IL-2 at 100 units per ml for both the overnight priming of effector cells (PBMCs, NK cells, or Splenocytes) and during the ADCC reaction. Miltenyi Biotec also has a modified IL-2 (IS) form that works well in assays.
Best Regards,
Omar
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