Dears, We are currently working with BAL cells and we have serious problems in detecting IL-4 and IL-10. We have performed intracellular staining for cytokines with reagents from Pharmingen. We tried various ways of stimulating T cells and the best one (and recommended by pharmingen) is stimulation with PMA/ionomycin. Great intercellular staining was observed with this activation methods for IFN-gamma, IL-2, we failed to detect any IL-4, or IL-10. The bottom line is that we are still struggling with this technique and have not found a solution to measure il-4 and Il-10.
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