Hello Wei

I work in macrophage polarization. Your experience is posible and I know about it. M1 and M2 markers dependent on type of cell and stimulus too. For this reason there are many candidates related with makers of polarization. Let me give you an advice, first choice a 3 o 4 genes related to M1 or M2 profile and test by PCR reaction. Then, choice 2 genes with the higher differential expression to M1 and M2 and buy their antibodies to test.

Best regards

This is not unusual. The classical way of defining macrophages is undergoing rethinking. There are many different macrophage phenotypes, not just M1/M2. Additionally, macrophages are highly plastic and can switch between these phenotypes depending on a number of factors (like stimulus, extend/time of treatment etc.). Even the best M1/M2 macrophage may not express all the markers that are used to define them.

I am attaching a couple links that might help you decide what kind of macrophage your antibody is inducing. If your macrophage phenotype does not fit into any of these boxes, you can just describe what they express and what they do not. You could explain the phenotype by looking at the mechanisms by which your antibody is inducing this phenotype (i.e. signaling pathways). That should be sufficient for scientists. It is ok for them to not have a name and to not fit into boxes that no longer make sense.

https://www.promocell.com/cells-in-action/macrophage-plasticity-black-white/

Article Macrophage Activation and Polarization: Nomenclature and Exp...

Thank you, it's useful answer. I use BV2 cell( one kind of microglia). I'm thinking there are two steps in macrophage function. First step is activation and second step polarization. Whether something inhibit the activation step will cause both M1 and M2 decrease? Because it keep the cell stay in monocyte phenotype. Oscar Rafael Escate Chávez Julie Joseph

I haven’t worked with microglial cells and so can’t be of much help. You may be able to find literature on what markers are upregulated/downregulated during maturation/differentiation of BV2 cells to macrophages. If you are worried that your cells are not getting activated and differentiating to macrophages, look at the expression of activation/differentiation markers. You could stain the cells with antibodies against these markers and then do flow cytometry.