I am trying to stain cytokine IL-4 in dendritic cells and Th2 lymphocytes recently. I am using a standard intracellular cytokine staining (ICS) protocol. Briefly, cells were stimulated (10ng/ml IL-4 for Th2 induction) and gave BFA blocking 4~5 hours before being collected for ICS analysis. anti-IL4 (1/200 dilution) antibody was used to staining Th2 cells (IL-4+ CD4+T cells). However, I barely detected any IL-4 expression in the culture system. Is there any specific tips I need to know for IL-4 cytokine staining?
Hi Qingrong,
IL4 staining can be tricky sometimes, especially because of its low amount. The blood sample preparation, the type and amount of Anticoagulant can be important step. The steps such as dendritic cells isolation as well as IL4 antibody amount are important. Finally there are especial IL4 kits in market with high sensitivity.
Thank you Behnam for your reply. I will try to check the IL4 kits then.
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