Hello - I'm working with stem cell derived microglia. When adding to 3D cultures I'm not getting a signal for TMEM119, which based on literature could be due to the immaturity of the cells (where TMEM is expressed postnatally). IBA1 is highly expressed in the cells.
Is there any other markers that would reliably determine if these cells indeed are microglia or if they're still just macrophages? (ie. a microglia specific marker to determine IBA+/microglia marker+ and IBA+/microglia marker-). Literature seems to be a little all over the board on this topic.
Have you had luck with other antibodies?
Thank you so much for the help!
Definitely a tricky question to get to the bottom of--- I haven't had much success with TMEM119 either, but you may want to try P2RY12 as a reliable microglial marker. It is often described as being specific for microglia (not peripheral macrophages). Our lab has had success with this clone: https://www.biolegend.com/en-us/products/purified-anti-p2ry12-antibody-14063
Good luck!
Elliot
Are you working on human or mouse cells?
As Elliot already mentioned P2RY12 is a valuable marker for microglia (the antibody that he mentioned works also great in our hands, but it is specific for mouse microglia). Another marker that distinguishes microglia from macrophages is Siglec H (again, murine system).
Commonly, also CX3CR1 is used for both human and murine microglia, but since you're working with stem cell-derived cultures, I'm not sure, whether this may work (don't know if your stem stells or intermediates may express CX3CR1).
So, try at least P2RY12 or (if in the murine system) Siglec H.
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