I'm optimising the PSD-95 antibody 7E3 (Santa Cruz Biotechnology) to stain some 30um brain slices (resectioned from 300um). The current protocol I'm using has not been very successful in that I'm not seeing as many puncta I would expect - literally just a few dots of PSD-95 per slice! I've used dilution factors of 1:50, 1:100, 1:250 and 1:500. I expected 1:500 would be more than enough since PSD-95 is such an abundant protein...
Has anyone had any success in doing this and is willing to share their methods? :)
Janine Dovey
Michel K Herde Thanks for that paper!
I'm fixing in 4% PFA and then putting the slices in 30% sucrose and letting them sink before resectioning.
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