M1 Macrophage Flow Cytometry Panel

Multicolor flow cytometry panel to identify M1 Macrophages. Monocytes were isolated from PBMCs via adherence depletion for 5 hours. Non-adherent cells were removed. Adherent cells were incubated for 6 days in C3 +10% huAB media with 50 ng/ml Recombinant Human GM-CSF (215-GM), media and cytokines are refreshed on day 3. For M1 polarization, 24 hours prior to staining, cells are incubated with 50 ng/ml human GM-CSF (215-GM), 50 ng/ml Recombinant Human IFN-gamma (285-IF), and 50 ng/ml LPS. Cells were stained with Anti-human CD3 Alexa Fluor ® 405, CD20 Alexa Fluor 405, CD56 Alexa Fluor 700, CD86 FITC, HLA-DR PerCP, CD11c PE,and CD38 APC. Cells were previously gated on Live CD3-CD20-CD56-. Find Staining Protocol Here.