I activate the CD8-T cell and perform ELISA for IL-2, PFP but the value of negative ctrl and ctrl are same, it means the cells are not activated. what is the specific procedure to know about the activation confirmation.
For a quick check, if your activation protocol works use anti-CD3 + anti-CD28 or PMA and ionomycin as positive control and if absolutely necessary assess CD69 (early activation markers for T cells) on the cell surface.
For more details, please see:
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Michael Förster let me try. before I added anti-CD3 + anti-CD28, but after performing ELISA, the IL-2, TNF-alpha, and IFN-gamma in negative control and control are the same, which means the cell did not activate or activate but didn't stimulated. let me try PMA and ionomycin. thank you for your response. hope it will work. thanks