Hey everybody,
does anyone have experience with kidney stainings (podocin) in embryos at 72 hpf? Wen want to image preferably the whole embryo without cutting afterwards.
I'm really new to this topic and am all alone with this in my group..I'm trying Dent's fixative+ MetOH/PBST rehydration and antibody staining as mentioned by IA Drummond in the Zebrafish book at the moment and plan to also go for 4/2% FA in PEM + 3%sucrose or PBS afterwards as the podocin-AB is mostly used with this combination.
What are your ideas for imaging afterwards ?
Thanks for your help!
You could TRY with fluorescent secondary Ab and perform confocal scanning of whole-mount embryos/larvae.
Maybe you have to specifically arrange a permeabilization step.
Anyway, here there are two excellent useful references (I guess you already checked literature...) for optimizing protocols.
https://link.springer.com/protocol/10.1385%2F1-59259-678-9%3A77
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3896505/
Hope this can help you.
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