when dc cells incubate with tumor lysate, then co-cultured with CD8+T cells.
And the tumor lysate to act as an antigen, there has two methods to get the tumor lysate: one is using several cycles of frozen-thaw; another is using lysis buffer. and should i add protease inhibitors cocktail and pmsf when making tumor lysates?
Hello Jiangnan He
I would recommend using several cycles of freeze-thaw to obtain tumor lysates. If you maintain the temperature below 4 degree C (on ice) while preparing the tumor lysates, the protease activity should be negligible. Use of protease inhibitors cocktail and pmsf when making tumor lysate could raise many doubts about the quality of tumour lysate, and you may not get the required results namely, activation of T cells.
Best.
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