Westerns-blotting-BSA

[Robert Hindges]

In article <Pine.SOL.3.91.960822141048.1166A-100000 at isnet>, Steve
<smiles at bgsm.edu> wrote:

> Hey all!
> 
> I'm doing a normal western blot, but our polyclonal antibodies seem to be 
> binding to many of the E. Coli proteins. It is possible that the 
> antibodies were raised to our protein, as well as any bacterial proteins 
> present in the blood during an infection in the rabbit.
> 
> Is there any protocol to add cell extract to the BSA blocking buffer, or 
> the primary antibodies, which will prevent all this binding?
> 
> Or is there a better way to block?
> 
> Thanks.
> 
> STEVE


hey Steve

I wonder if you see the background already with the pre-immune antibodies.
sometimes the rabbits show a background already before immunization. in my
case I had to test several rabbits, until I chose one for immunization. it
is a little bit more work, but sometimes it is worth it. 

robert.