[Histonet] HRP conjugation

[Donin, Nick (NIH/NCI)]

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 your help, any thoughts or suggestions are greatly
appreciated. 

 

Nick Donin

CRTA

Neuro-Oncology Branch

National Cancer Institute

National Institutes of Health

9000 Rockville Pike

Building 35, Room 2B-203

Bethesda, MD 20892