[Histonet] RE: HRP conjugation

[C.M. van der Loos]

   Hello Nick,

   I   am   wondering   if   you  could use  a  Mouse-on-mouse  detection
   system for this  purpose. Try  Vector MOM kit or Dako ARKit. These kit
   are definitely much more simpler than "true" HRP-conjugation.

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



   Date: Fri, 18 Nov 2005 13:14:30 -0500
   From: "Donin, Nick \(NIH/NCI\)" <doninn <@t> mail.nih.gov>
   Subject: [Histonet] HRP conjugation
   To: <histonet <@t> lists.utsouthwestern.edu>
   Hello Everyone,
   This isn't a Histology question, but it relates to antibodies, so I
   thought someone could help me.  I'm doing a western blot, and I'm
   running the protein from mice tumors.  I'm using a mouse primary
   antibody  to  stain  for a particular antigen, but the problem is that
   when
   I apply anti-mouse secondary, it stains everything on the blot
   (presumably because the tumor tissue is rich in mouse IgG antibodies).
   I thought I could get around this by conjugating my primary antibody
   directly to H.R.Peroxidase, thus eliminating the need for a secondary.
   My first question is:
   1. Does this strategy even make sense?
   2. Does anyone have a technique or product for carrying out this
   conjugation?

   Thanks for all yo! ur help,