[Histonet] RE: Background staining: mouse on mouse Broad spectrum

[C.M. van der Loos]

   Dear Cathy,

   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
   ([1]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
   whatever.

   This  doesn't  answer  your  questions, but might solve the background
   staining problem!

   Hope this works for you.

   Chris van der Loos, PhD
   Dept. of Pathology
   Academical Medical Center M2-230
   Meibergdreef 9
   NL-1105 AZ Amsterdam
   The Netherlands

   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
   kit
   To: histonet <@t> lists.utsouthwestern.edu
   Dear All,
   I  really  need  some  advice  regarding an ongoing intense background
   problem.
   We use FFP sections of mouse liver.
   We  are  using  an immuno kit called Zymed Histomouse Max Kit (It is a
   HRP-
   DAB one).
   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
   to be
   used for mouse, rabbit, rat and guinea pig primaries.
   The  secondary  antibody  is not completely explained in the kit but I
   think
   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
   it
   actually is.
   Question: If the second! ary ab is

References

   1. http://probes.invitrogen.com/products/zenon