What antibody are you trying to use?
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Today's Topics:
1. Help with Biocare polymer detection kit!! (Jennifer Campbell)
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Message: 1
Date: Sat, 31 Jul 2010 12:03:39 -0700
From: "Jennifer Campbell" <jcampb...@vdxpathology.com>
Subject: [Histonet] Help with Biocare polymer detection kit!!
To: <histonet@lists.utsouthwestern.edu>
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I have been trying to get the Mach 1 polymer kit from Biocare to work on my K9
and feline tissue and I am getting terrible nonspecific staining in my positive
and negative controls. It appears to be sticking to something in the glands of
the intestinal epithelia. I am also getting nonspecific staining in my LN
tissue.
I decided to switch over to polymer detection kit because I had been dealing
with the issue of endogenous biotin when using the streptavidin biotin-based
detection system, and now I am getting this other nonspecific staining!
I have tried doing runs eliminating my protein block and eliminating the
negative control and primary antibody step altogether (to see if maybe the
protein block or the diluent used in my negative control and antibody dilutions
are causing the problem). I have also reduced the incubation time of both the
probe and polymer from 15 min and 30 min, respectively, to 10 min each. I have
tried cutting down my antigen retrieval time substantially as well.
I'm not sure what else could be causing this. Any suggestions??
Thanks in advance,
Jennifer Campbell
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