[Histonet] Immunofluorescence in paraffin sections
I am doing immunofluorescence in paraffin sections. I have some difficulties in it:
1: For pretreatment of tissue sections, I plan to incubate sections with trypsin solution, but I do not know :what kind of concentration should I choose ? (0.01% trypsin, 0.05% or o.1%)how long does it should last? (10min,20min,30 min)
2: Recently, I read a protocol about double immunofluorescence, it shows :Antibodies derived from different animal can be mixed and incubated as a cocktail (example: rabbit anti-A and mouse anti-B). The same is valid for secondary antibodies (example: goat anti-rabbit Texas Red conjugated and goat anti-mouse fluorescein conjugated). but If secondary antibodies cross-react, what should I do? My friend told me that I could use depletion method. but I am not sure.
Can anybody do me a favor?
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