Need a lot more details... What clone of anti-IL-4, IL-10? What fluorochromes for each antibody? Staining scheme? duration of activation? activation conditions (media, concentrations)? What fixation and perm reagents? BAL in what patient population or for that matter what species? What is your negative control? What is your positive control? Have you got this technique working in PBMC? How do you process BAL? Calman > _______________________ > Calman Prussin > Laboratory of Allergic Diseases > NIAID/ National Institutes of Health > > ---------- > From: Karatza > Sent: Wednesday, November 7, 2001 8:57 > To: Cytometry Mailing List > Subject: intracellular IL-4 and IL-10 in BAL > > 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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