in my study about meassuring the production titer of IFN gamma and IL-10, in my previous experiment the control well (PBMC culture + Medium) show me the number of IFN gamma and IL-10, but now im lil bit confused how to know the normal titer of IFN gamma and IL10 from PBMC culture?
Hi
I have done IFN g ELISpot assay after 14 days culture of my effector : target cells. I normally use untouched cells/unstimulated cells for control. I will take the spots obtained in the unstimulated control as a base line and normalize the spot by subtracting the spot count obtained in my unstimulated in stimulated spots.
Are you stimulating the PBMCs and then measuring the IFN-gamma and IL-10 Levels
Usually the Unstimulated cells will secrete lower levels of these cytokines below 10pg/ml
@Dhandapani and @Kamran Haq Thank you so much for your suggestion
@Asif Amin Dar: yeap... the unstimulated cell secrete the low level... so it's normal right?
Yes and on stimulation the levels will increase depending on the stimulation
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