Hello flowers, I have the following problem. I will make some experiments with measuring intracellular cytokines by flowcytometer. So I try to see how this method works. The protocol is on BD. We are using PMA and I for activation. First, the problem was that we had not enough intracellular cytokines after the activation. I found that using diluted PMA stock that has been at -70o for one year does not work properly. It is not the case with Ionomycin. I just compared the new and the old solutions and my conclusion is that PMA is very sensitive substance compared to Ionomycin. So I made the intracellular protocol with new solutions and again I have a problem. I repeat the protocol twice (in 2 successive days) using blood samples from one subject. In one of the measurement all intr. Cytokines were 1.5-2.5 times lower than the other one. In one other experiment 6 out of 20 subjects showed quite different intracellular cytokines (variation within a subject 1.5-2.5 times) when 2 blood samples were obtained from a subject in interval of one week. My question is: Is it possible to have such (within a subject) variation in intracellular cytokines in whole peripheral blood? Or the problem is activation. Thank you in advance Stoyan Klinische Forschergruppe Neuroendokrinologie Medizinische Universität Lübeck Ratzeburger Allee 160 Haus 23a 23538 Lübeck Tel.: +49 (451) 500-3644 Fax.: +49 (451) 500-3640
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