[Histonet] RE: Background staining: mouse on mouse Broad spectrum
C.M. van der Loos
c.m.vanderloos <@t> amc.uva.nl
Wed Jul 27 02:54:33 CDT 2005
From your mail I understand this Zymed Histomouse Max Kit isn't
working for you. I heard from a colleague here in the hospital this
kit wasn't working background-free for her on rat tissue either. I
realized the Dakocytomation ARKit (based on streptavidin-biotin) isn't
appropriate here as well since you are dealing with liver full of
non-blockable endogenous biotin. The best alternative for you is
perhaps a Zenon kit
(http://probes.invitrogen.com/products/zenon). Like with the
ARKit the Zenon kit works with an in vitro labeling of the primary
antibody with a labeled Fab-fragment. Kits are available with many
fluorochromes as label, including FITC. This allows further enzymatic
visualization via Rabbit anti-FITC and goat anti-rabbit/HRP or
This doesn't answer your questions, but might solve the background
Hope this works for you.
Chris van der Loos, PhD
Dept. of Pathology
Academical Medical Center M2-230
NL-1105 AZ Amsterdam
phone: +31 20 5665631
fax: +31 20 6960389
Date: Tue, 26 Jul 2005 18:44:09 +1000
From: Cathy Malcontenti-Wilson <cmalc <@t> unimelb.edu.au>
Subject: [Histonet] Background staining: mouse on mouse Broad spectrum
To: histonet <@t> lists.utsouthwestern.edu
I really need some advice regarding an ongoing intense background
We use FFP sections of mouse liver.
We are using an immuno kit called Zymed Histomouse Max Kit (It is a
Our primary ab is mouse anti human VEGF (1:1000).
The kit is called a BROAD SPECTRUM kit because it is meant to be able
used for mouse, rabbit, rat and guinea pig primaries.
The secondary antibody is not completely explained in the kit but I
it is of goat host species.
Question: If it is broad spectrum does this mean that the secondary
antibody is a mixture of different antibodies?
The kit contains its own non specific block, with no details of what
Question: If the second! ary ab is
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