I will study intracellular cytokine (IFN-g, IL-4) production in T cells for clinical trials. The plan is to collect and freeze PBMC in different time points, then thaw the cells and stimulate the cells with antigen on same day for cytokine detection. BD offers FastImmune intracellular cytokine kits (IFN-g or IL-4/CD69/CD4/CD3) for whole blood. Based on the information from its FastImmune CFC Handbook, it seems the FastImmune kit can also be used on frozen PBMC. I would like to know if you have the (postive or negative) experience in using the kits on frozen PBMC, and experience in using the kits for on whole blood (stimulation/BFA, lyse/fix, permeablization, staining on one day) then freeze the cells at -80C for later analysis. Thank you, Dianna Wu BMS
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