[Histonet] Annexin V IHC staining in mouse xenograft

[Amos Brooks]

Hi,
    Have you considered running the antibody through some antibody
purification beads. It isn't really a common histolgy thing to do, but the
process isn't terribly difficult. It really sounds like a dirty antibody is
at fault. It will probably result in a cleaner and more concentrated
antibody. There are some commercially available kits to do this. I believe
Invitrogen carries one and I'm almost sure Chemicon does too.

Amos


Message: 1
Date: Tue, 11 Oct 2011 17:11:38 +0000
From: <awatanabe <@t> tgen.org>
Subject: [Histonet] Annexin V IHC staining in mouse xenograft
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <CAB9C6E3.925E%awatanabe <@t> tgen.org>
Content-Type: text/plain; charset="us-ascii"

Hi histonetters,
I need some help.  I am running an antibody from Biosensis called Annexin V
in mouse xenograft brain samples with human glioma tumors treated and
non-treated.  I have incredible background that I can not get rid of any of
my samples.  I have used a protein block, rodent tissue block, post
fixation, short antibody incubation time, short antigen retrieval time, very
dilute antibody, polymer detection and every combination in between.  I'm
looking to see if anyone knows anything else to try.  I'm trying to
rearrange the protocol to put steps in different places to see if that
works.  At this point I'm not sure what else to try so any ideas would be
great.

Aprill Watanabe, B.S.
Research Associate II