Why PD-1 expression go higher if I do nothing to them only to put them outside on the medium for 24 or 48 hr?
I think it should be stimulate ..then the PD-1 will express higher
Hi Yunuo,
What cells are you working with splenocytes? Are you using Flow cytometry? Are you blocking the Fc receptors in your sample prior to staining? Do you run isotope controls when you do your staining?
Sorry I should talk about the background more.I use the normal PBMC and stimulate them with CD3/CD2 8beads ,also use the pre-stimulate PBMC as control.However I found out when I use the Flow cytometry to see the expression of PD-1
PD-1 in pre-stimulated samples vs stimulated samples ,the expression is almost the same ,so I wondering what's the problem of it ..is that because I put the PBMC outside in RPMI CM for 24 hr and 48 hr cause that ,or because I use the frozen sample cause that ,I have no idea..I don't use the isotype ..only use the unstained as a control
I think you should try blocking Fc receptor using anti-CD16/32 blocking and also add isotype controls.
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