Hi there, This is the first time that I am writing to you, a colleague gave me this email. I have some questions regarding the intracellular staining (CYTOKINES) on PBMC (comimg from Parasites infected patients), I would work in Kenya on PBMC fresh cells, but I was wondering if we could get the same data on fresh cells than on frozen cells, if the answer is yes, does somebody have a protocol which works perfectly . Ususally the simulus used for PBMC Is PMA Ionomycine, did somebody try to stimulate with a specific antigen (from the parasite by instance), and have a more specific respone, the PMA Iono is fine but up to me probably such strong that we could not detect small variations. The other question I have was on mouse splenocytes. I do some intracellular staining on parasite infected mice, and was wondering, again, in the mouse model, if somebody tried to stimulate with specific antigen rather than PMA+ionomycine and got some nice or bad results. I hope these questions are clear enough to be understood. Thank you very much in advance.
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