I am trying to stain drosophila third instar imaginal discs (wing) using
an anti-achaete antibody and HRP-DAB staining. However I either cannot
get any signal at all or too much background so that the staining
pattern can't be made out! This is trying different fixation methods. I
have tried 4% formaldehyde for various times and temperatures down to
1%. All of my reagents are ok so it must be my method. Can anyone
Do your staining kits use biotin at any step? I had that problem when I
used the Vector Elite kit, where you use a 2¡ ab-biotin/strepavidin-HRP
mix. When I used a 2¡ Ab conjugated to HRP, the background went away.
Hope that helps.