Re: amplification kit

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From:<> (Karen Larison) (by way of histonet)
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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
tyramides, but background is definitely a problem in frozen sections when I
use the
fluorescein tyramides.  Titrating the primary antibody doesn't help.  Oddly
if I use the fluorescein tyramide on whole embryos, I get gorgeous staining
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 <>
Subject:       Re: amplification kit
To:            Klazien Bosch <>

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 Tuomala
Anatomic Pathology
St.Joseph's Hospital
Hamilton, Ontario, Canada

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