M2 Macrophage Flow Cytometry Panel

Multicolor flow cytometry panel to identify M2 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 M-CSF (216-MC), media and cytokines are refreshed on day 3. For M2 polarization, 24 hours prior to staining, cells are incubated with 50 ng/ml human M-CSF (216-MC), 20 ng/ml Recombinant Human IL-4 (204-IL), and 20 ng/ml Recombinant Human IL-13 (213-ILB). Cells were stained with Anti-Human CD3 Alexa Fluor® 405, CD20 Alexa Fluor 405, CD56 Alexa Fluor 700, CD206 Alexa Fluor 488, CD163 PerCP, CD204 PE, and VEGF APC. Cells were previously gated on Live CD3-CD20-CD56-. Find Staining Protocol Here.