Hi, I want to evaluate stress oxidative markers in macrophages using western blot. I want to know an easy protocol to isolate macrophages from PBMCs, I already know a protocol to isolate them using ficoll, but after that I don't know what to do. I read about plastic adhesion. I was thinking about coating a T-75 flask with 2% gelatin. Then I'll plate PBMCs on the coated flask and wait 1 hour to wait monocytes to adhere (in this step I use RPMI 1640). For the differentiation, ¿it is ok to use RPMI 1640, GM-CSF 10ng/ml, and 10%FSB?. I really appreciate any suggestions.
Could you describe the characteristics of peripheral blood mononuclear cells to the audience?
PBMCs are composed of lymphocytes, monocytes and dendritic cells. They’re are isolated from peripheral blood by centrifugation, that removes red blood cells and granulocytes cells. PBMCs, mostly are cells in a resting state, with no effector functions.
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