Re: amplification kit

<< Previous Message | Next Message >>
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




<< Previous Message | Next Message >>