Hello, we are developing a protocol for combining FISH and IHC on Drosophila embryonic brain, using FISH to localize mRNA and IHC (2 antibodies) to mark neuropil and neural tracks. We thought about mixing anti-DIG antibodies with one of the IHC antibodies, but our usual protocol for IHC calls for incubation in antibody for one to two days at 4ºC, and the protocol I found for anti-DIG says to incubate for at least 4 hours at RT. So now we're planning to separate the two processes, but we don't know which one we should do first. Any protocol or suggestions will be appreciated!

If it helps, here are some of the materials we're using:

Anti-DIG-Fluorescein Fab Fragments from Roche

IHC blocking: 100ul normal goat serum + 900ul PBT

IHC protocol: PBT wash (2x20min) -> block (30min) -> 1º staining (O/N) -> PBT wash (3x20min) -> block (30min) -> 2º staining (O/N) -> PBT wash (30min)

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