Re: amplification kit
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From: | <larisonk@uoneuro.uoregon.edu> (Karen Larison) (by way of histonet) |
To: | histonet@histosearch.com |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
If I remember correctly, Mr Bosch was trying to use biotinylated tyramides
to amplify
signals in frozen sections. I haven't had any experience with the
biotinylated
tyramides, but background is definitely a problem in frozen sections when I
use the
fluorescein tyramides. Titrating the primary antibody doesn't help. Oddly
enough,
if I use the fluorescein tyramide on whole embryos, I get gorgeous staining
with
virtually no background. Is the substrate sticking to the cut membranes?
It's a
mystery to me!
Karen in Oregon
Date: Thu, 16 Dec 1999 12:43:58 -0500
From: Katri Tuomala <katri@istar.ca>
Subject: Re: amplification kit
To: Klazien Bosch <k.s.bosch@amc.uva.nl>
Cc: HistoNet@pathology.swmed.edu
Hi Klazien,
You may have received the answer to your inquiry already, but I have not
seen it. I have used this kit on paraffin sections with antibodies that
are normally not demonstrable in paraffins. I had to do a lot of
tweaking with the antigen retrieval and antibody dilutions, because of
the background staining. I have a feeling you may be successful with a
much higher primary antibody dilution. Do a wide titration first to get
closer to real titre and then narrow it down. Good luck!
Katri
Katri Tuomala
Anatomic Pathology
St.Joseph's Hospital
Hamilton, Ontario, Canada
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