[Histonet] Stripping antibodies
Amos Brooks
amosbrooks <@t> gmail.com
Sat Oct 29 12:40:22 CDT 2011
Hi,
Stripping sites are usually found in the more seedy areas of large
cities ... Oh wait you meant ... nevermind :-) If you developed the reaction
with DAB, you may be in a difficult position. DAB is a really strong
reaction. You might try a Mallory bleach to remove it. 0.3% sulfuric acid in
5% potassium premanganate for 5 min followed by 5% oxalic acid until it is
clear (usually a couple of minutes). Unfortunately, I am not sure what
effect this will have on the epitopes you are looking for, but this is
usually what is used to clean precipitated DAB from instruments it should
work on the tissue too.
If you use another chromogen such as AEC it is much easier (to many
people's dismay) to remove this end product. Using alkaline phosphatase will
be easy to remove the chromogens as well. It would also be very easy to do
this with fluorescent tags too. Just remove the coverslip and dip the slide
in ethanol and it will remove the colored end product. Then just start over
for the new antigen.
Good luck,
Amos
On Fri, Oct 28, 2011 at 11:50 AM, <histonet-request <@t> lists.utsouthwestern.edu
> wrote:
> Message: 2
> Date: Thu, 27 Oct 2011 10:30:40 -0700
> From: "Claire Weston" <cweston <@t> valasciences.com>
> Subject: [Histonet] Stripping antibodies
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <002401cc94ce$2619ca60$724d5f20$@com>
> Content-Type: text/plain; charset="us-ascii"
>
> Hi!
>
>
>
> Does anyone have experience in IHC stripping antibodies from a tissue
> section and re-probing with different antibodies? What is the best way to
> do that? If you could share your stripping protocol or experience I would
> appreciate it - thanks!
>
>
>
> Claire
>
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