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To: bionet-drosophila@net.bio.net
From: sa08383@odin.mdacc.tmc.edu (Kathleen Gajewski)
Newsgroups: bionet.drosophila
Subject: Re: Imaginal disc ab staining
Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre
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Date: Fri, 27 Jul 2001 21:42:00 +0100 (BST)
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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
help????


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.

						Kathleen

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