cytokines release (or intracellular) measurement

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cornillon
cornillon's picture
cytokines release (or intracellular) measurement

is it possible to mesure the cytokine release (like IFNg) of NKL after a cytotoxicity assay with target cells and antibodies, in vitro. And if it is possible, what is necessary (cytokines or other things) in the medium of the test. because I made it with murine NK ex-vivo (activation in vivo), and I don't know if you can do it with in vitro cultured cells and NKL particularly.

samm
samm's picture
Yes, you can do that.

Yes, you can do that.
There are two ways to go about it:
1) Bead conjugated antibodies detect extracellular cytokines secreted in the course of the assay. These assays also allow for "multiplexing" - by varying the size of beads, and colors, as many as 8 cytokines can be measured in a standard 5-parameter FACS machine.
2) Intracellular staining (in the presence of BFA or monensin) actually takes 2-6 hr extended "snapshots" of cytokine production.

Both these assays can be semi-quantitative and are analyzed by flow cytometry.

cornillon
cornillon's picture
thank you. But I have nothing

thank you. But I have nothing else to add to the medium (like IL-2 or something else)? If I can do that with my cells only it would be great!! I hope it would be ok!!
Thanks again.