I’m looking at axons from mouse and human DRG in vitro. The standard stain is TUBB3. I’m using the Tuj-1 clone from Biolegend (formerly Covaris), which is the gold-standard as far as I can tell.
I’m using a standard ICC protocol (1% BSA, 0.1% triton in PBS) and 1:500 antibody. In mouse there is faint satellite glia/fibroblast? staining and in human it seems that it is very strong. In the mouse, there are clearly axons, but I think also some non-neuronal cells.
Has anyone else seen this?
Also, what are some good alternatives for specific axon staining in vitro?
In the past, we’ve used PGP9.5 to look at neuronal axons in the skin (fixed sections, not cell culture). We didn’t see any non-neuronal staining in that situation, but we also have not looked in the DRG itself. I think PGP9.5 is supposed to be somewhat neuronal-specific, although I can’t confirm that from direct experience. Since we were looking in the skin, we didn’t see any cell bodies, but if you’re looking at cell culture, I’m sure you’d get somatic staining as well.
