RE: Normal isoform PrP
|From:||Cynthia Favara <firstname.lastname@example.org>|
3F4 reacts with both the normal and abnormal forms of PrP. The
abnormal form of PrP needs to be unfolded in order for the epitope to be
available. I have done autoclaving with water and with 0.1mM HCl and my
experience is that very little staining of abnormal PrP is seen with water
and this is enhanced with 0.1mM HCl. I do not use a biotinylated secondary
but a HRP conjugated secondary and you may have a better signal because of
that additional layer.
Rocky Mountain Laboratories
903 S. 4th Street
Hamilton, MT 59840
From: Tina Cardamone [mailto:email@example.com]
Sent: Monday, November 13, 2000 12:01 AM
Subject: Normal isoform PrP
I have a quick question regarding CJD brains and immunohistochemistry.
I stain with the Senetek monoclonal antibody 3F4.
It labels normal isoform PrPc and the infectious form PrPsc.
I autoclave the sections in water for 30 mins at 132 deg. cel.
I block endogenous peroxidase with hydrogen peroxide and block non specific
binding with the appropriate serum and then stain using the Avidin Biotin
method.I get great results.
My question is, am I blocking or digesting away the normal PrP (PrPc) with
the autoclaving I do or am I staining for both ie PrPc and PrPsc. Is there
something else I should be doing to eliminate PrPc.
I am looking forward to hearing from you.
Thank You in advance.
Uni of Melbourne
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