I would like to phenotype M1 and M2 microglia in some specific brain region of the mouse brain. I am currently looking specifically at antibodies against Arg-1, iNOS and CD206... Could anyone recommend good antibodies ie specific, great staining, correct dilution range for Immunofluorescence staining of mouse brain section?
If you can, I would be grateful if you can provide Company, Catalogue number and any tips for IHC with those antibodies ;-)
I have been looking at the literatures but unfortunately I got caught in the indefinite loop of "as previously described [citation]"....or the M&M miss information either about provider but mostly missing catalogue number 🤪
Thanks a lot!
Dear Elodie!
We have done several sets of IHC on 4µm mouse brain sections using the Leica Bond III system (Leica Biosystems Newcastle Ltd, UK).
The paraffin-embedded tissue blocks were chilled on ice and sliced on a Leica (Leica Biosystems, Nussloch, Germany) RM2235 manual rotary microtome at a thickness of 4 μm. The sections were mounted onto gelatin-coated slides, dried overnight at room temperature, and stored at 4 °C inside storage boxes.
Tissues were pretreated with epitope-retrieval solutions (ER, Leica Biosystems Newcastle Ltd, UK) followed by 30 minutes incubation with primary antibodies.
We used ARGI (GeneTex, GTX113131), CD68 (Abcam, # ab125212) and ARGII (GeneTex, #GTX104036) Abs with dilutions 1:500, 1:200, and 1:600, respectively. ER/time 2/20 (min)
Of note, IBA1 Ab was used as a standard marker for microglia.
The Leica Refine-HRP kit (Leica Biosystems Newcastle Ltd, UK) used for detection and counterstain with Hematoxylin.
Best regards
https://link.springer.com/article/10.1007/s13311-018-0669-5
https://www.mdpi.com/1422-0067/20/18/4616/htm
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