Hi I am measuring intracellular Cytokines in Monocytes, but I am getting bad results with IL-10, all other Cytokines work great with my assay. I get about 2-5 % positve Cells after 6h Stimulation with LPS (10nM), but even these results are very unstable. With PMA and Ionomycin the results vary much more than with LPS. Mostly I can not detect IL-10 at all. So I got some questions about this matter. 1. Which Stimulus works best to get IL-10 ? and which Concentration you use for Stimulation ? 2. Which point of time is the best to measure IL-10 ? 3. How much positive Cells I have to expect at that point of time and with which Stimulus ? 3. Which Golgi-Inhibitor is the best to measure it ? Monensin or Brefelidin A ? what concentrations you use? 4. Is it possible that i loose my monocytes, which are IL-10 postive through cell adherence in my Tubes? Thanks in advance Philipp Philipp K. Röntgen Department of Thoracic Surgery MLU-Halle Heinrich Damarow Str.1 06120 Halle Germany
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