When I do NK or iNKT cytotoxicity with Jurkat cells or attachment cells, found these issues:
1. iNKT cells and attachment cells are co-culture for 15h, then add protein transport inhibitor cocktail (Brefeldin A und Monensin, 500X, eBioscience) and CD107a antibody (Biolegend,328608) for 7h. Then to do cells washing and flow assay. We found the iNKT cells almost express CD107a and suppose maybe some issues with the testing.
2. In order to solve this issues, we stain iNKT cells or Jurkat cells with CD107a seperately without add protein transport inhibitor cocktail. Then to do flow assay and the results are the same as the first one testing's.
Could some give me some more suggestions about this assay?
Very apprecaite for all of you.
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