Hi all,
I'm trying to perform a whole mount double ISH on 48 dpf zebrafish.
I've followed standard protocols using anti-DIG with fast red (sigma tablets), and fluorescein with TSA (Alexa Fluor™ 488 Tyramide Reagent, Thermo Fisher). Unfortunately it isn't working (no color after staining). In parallel I've performed regular ISH as a control, showing that DM purple does stain and it worked.
I would like to hear recommendations or protocols using these materials.