Macrophages and myeloid dendritic cells can be proinflammatory or express suppressive, anti-inflammatory characteristics. One characteristic that is part of the proinflammatory phenotype is the expression of inducible nitric oxide synthetase (iNOS), which can be evaluated by measuring production of nitric oxide using our all-inclusive Nitric Oxide Assay Kit.
This easy-to-use NO Kit can give rapid information about the differentiation state of myeloid cell types and guide experiments to the desired conditions. If you want to create M1 or M2 macrophages, this assay kit is one of the simplest ways to test the success of your protocol to skew the phenotype to M1.
Nitric Oxide Kit in Action
In testing our NO Kit, we thawed our proinflammatory mouse dendritic cells (DC) and placed them in culture with LPS to stimulate the nitric oxide production. We then collected culture medium after 24, 48 and 72 hours and measured the amount of nitrite in the medium using our NO Kit.
As shown in the figure, NO was produced following stimulation with LPS and increased each day.
Note that our mouse DC were differentiated from bone marrow cells using GM-CSF, resulting in proinflammatory DC. Bone marrow-derived macrophages did not produce NO unless pretreated with interferon gamma prior to stimulation with LPS.